quantitect sybr green pcr kit  (Qiagen)

 
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    QuantiTect SYBR Green PCR Kit
    Description:
    For real time PCR and two step RT PCR using SYBR Green Kit contents Qiagen QuantiTect SYBR Green PCR Kit 40 x 50L rxns cDNA and DNA Sample Real time PCR and two step RT PCR Reaction With ROX Ideal for Real time Quantification of Genomic DNA or cDNA Targets For Real time PCR and Two step RT PCR Using SYBR Green Includes 1mL 2x QuantiTect SYBR Green PCR Master Mix 2mL RNase free Water Benefits High PCR specificity with integrated hot start Reliable quantification of low abundance transcripts Accurate quantification over several logs of template Available with or without uracil N glycosylase UNG No need to optimize reaction and cycling conditio
    Catalog Number:
    204141
    Price:
    127
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    QuantiTect SYBR Green PCR Kits
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    Structured Review

    Qiagen quantitect sybr green pcr kit
    QuantiTect SYBR Green PCR Kit
    For real time PCR and two step RT PCR using SYBR Green Kit contents Qiagen QuantiTect SYBR Green PCR Kit 40 x 50L rxns cDNA and DNA Sample Real time PCR and two step RT PCR Reaction With ROX Ideal for Real time Quantification of Genomic DNA or cDNA Targets For Real time PCR and Two step RT PCR Using SYBR Green Includes 1mL 2x QuantiTect SYBR Green PCR Master Mix 2mL RNase free Water Benefits High PCR specificity with integrated hot start Reliable quantification of low abundance transcripts Accurate quantification over several logs of template Available with or without uracil N glycosylase UNG No need to optimize reaction and cycling conditio
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    1) Product Images from "Zinc oxide nanoparticles selectively induce apoptosis in human cancer cells through reactive oxygen species"

    Article Title: Zinc oxide nanoparticles selectively induce apoptosis in human cancer cells through reactive oxygen species

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S29129

    Quantitative real-time polymerase chain reaction analysis of mRNA levels of apoptotic genes in human lung cancer (HepG2) cells treated with zinc oxide nanoparticles (ZnO NPs) at the concentration of 15 μg/mL for 24 hours. Quantitative real-time polymerase chain reaction was performed by QuantiTect SYBR Green PCR kit using an ABI PRISM 7900HT sequence detection system. The β-actin was used as the internal control to normalize the data. ZnO NP-induced alterations in mRNA levels are expressed in relative quantity compared with those for the respective unexposed control cells. ( A ) p53, ( B ) bax, ( C ) bcl-2, and ( D ) caspase-3. Data represented are mean ± standard deviation of three identical experiments made in triplicate. Note: *Statistically significant difference as compared with the controls ( P
    Figure Legend Snippet: Quantitative real-time polymerase chain reaction analysis of mRNA levels of apoptotic genes in human lung cancer (HepG2) cells treated with zinc oxide nanoparticles (ZnO NPs) at the concentration of 15 μg/mL for 24 hours. Quantitative real-time polymerase chain reaction was performed by QuantiTect SYBR Green PCR kit using an ABI PRISM 7900HT sequence detection system. The β-actin was used as the internal control to normalize the data. ZnO NP-induced alterations in mRNA levels are expressed in relative quantity compared with those for the respective unexposed control cells. ( A ) p53, ( B ) bax, ( C ) bcl-2, and ( D ) caspase-3. Data represented are mean ± standard deviation of three identical experiments made in triplicate. Note: *Statistically significant difference as compared with the controls ( P

    Techniques Used: Real-time Polymerase Chain Reaction, Concentration Assay, SYBR Green Assay, Polymerase Chain Reaction, Sequencing, Standard Deviation

    2) Product Images from "Fast detection of Noroviruses using a real-time PCR assay and automated sample preparation"

    Article Title: Fast detection of Noroviruses using a real-time PCR assay and automated sample preparation

    Journal: BMC Infectious Diseases

    doi: 10.1186/1471-2334-4-15

    T m analysis of real-time PCR with SYBR Green as fluorescent dye. T m analysis allows the differentiation of specific PCR product (amplicon) from non-specific amplification products, such as primer-dimers. Samples: 1 to 7, patient samples (stool); 8, negative control (water); 9, positive control (NoV positive stool). Amplified products from NoV template RNA can be reliably distinguished from non-specific products by different melting points. The melting temperature for the positive samples was 79.0 +/- 0.5°C. Non-specific products melt at lower temperatures.
    Figure Legend Snippet: T m analysis of real-time PCR with SYBR Green as fluorescent dye. T m analysis allows the differentiation of specific PCR product (amplicon) from non-specific amplification products, such as primer-dimers. Samples: 1 to 7, patient samples (stool); 8, negative control (water); 9, positive control (NoV positive stool). Amplified products from NoV template RNA can be reliably distinguished from non-specific products by different melting points. The melting temperature for the positive samples was 79.0 +/- 0.5°C. Non-specific products melt at lower temperatures.

    Techniques Used: Real-time Polymerase Chain Reaction, SYBR Green Assay, Polymerase Chain Reaction, Amplification, Negative Control, Positive Control

    3) Product Images from "Friend of GATA suppresses the GATA-induced transcription of hepcidin in hepatocytes through a GATA-regulatory element in the HAMP promoter"

    Article Title: Friend of GATA suppresses the GATA-induced transcription of hepcidin in hepatocytes through a GATA-regulatory element in the HAMP promoter

    Journal: Journal of molecular endocrinology

    doi: 10.1530/JME-11-0060

    The expression of GATA (2, 3, 4, and 6; panels A–D) and FOG (1 and 2; panels E and F) in the human liver and hepatoma cell lines. Total RNA from hepatocytes was isolated using TRizol (Invitrogen Canada). An aliquot of the total RNA (2 μg) was used as a template to synthesize the first-strand cDNA using the Omniscript reverse transcriptase-PCR system (Qiagen), 5 μl of diluted cDNA (1:6) was then used as a template in the RT-PCRs. Quantification of gene expression was performed by a Rotor Gene 3000 Real-time DNA Detection System (Montreal Biotech, Kirkland, QC, Canada) with QuantiTect SYBRGreen I PCR kits (Qiagen). Amplifications were performed in duplicate using at least three different preparations of first-strand cDNAs for each of the three different RNA extractions while adult human liver total RNA was purchased from Clontech Laboratories, Inc. Expression levels for GATA (dark gray bars) and FOG (light gray bars) were normalized to the housekeeping gene β-actin. The results are presented as mean (±S.E.M.) of the relative mRNA expression. A different superscript indicates a statistically significant difference between means ( a–l P
    Figure Legend Snippet: The expression of GATA (2, 3, 4, and 6; panels A–D) and FOG (1 and 2; panels E and F) in the human liver and hepatoma cell lines. Total RNA from hepatocytes was isolated using TRizol (Invitrogen Canada). An aliquot of the total RNA (2 μg) was used as a template to synthesize the first-strand cDNA using the Omniscript reverse transcriptase-PCR system (Qiagen), 5 μl of diluted cDNA (1:6) was then used as a template in the RT-PCRs. Quantification of gene expression was performed by a Rotor Gene 3000 Real-time DNA Detection System (Montreal Biotech, Kirkland, QC, Canada) with QuantiTect SYBRGreen I PCR kits (Qiagen). Amplifications were performed in duplicate using at least three different preparations of first-strand cDNAs for each of the three different RNA extractions while adult human liver total RNA was purchased from Clontech Laboratories, Inc. Expression levels for GATA (dark gray bars) and FOG (light gray bars) were normalized to the housekeeping gene β-actin. The results are presented as mean (±S.E.M.) of the relative mRNA expression. A different superscript indicates a statistically significant difference between means ( a–l P

    Techniques Used: Expressing, Isolation, Polymerase Chain Reaction

    4) Product Images from "The Identification and Characteristics of Immune-Related MicroRNAs in Haemocytes of Oyster Crassostrea gigas"

    Article Title: The Identification and Characteristics of Immune-Related MicroRNAs in Haemocytes of Oyster Crassostrea gigas

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0088397

    The expression of eight miRNAs detected by SYBR Green real-time PCR in oyster haemocytes after bacteria challenge and heat stress, including cgi-miR-8 (A), cgi-miR-12 (B), cgi-miR-100 (C), cgi-miR-125 (D), cgi-miR-1984 (E), scaffold631_909 (F), scaffold42648_5080 (G) and scaffold1599_5643 (H). 5S gene was used as an internal control to calibrate the cDNA template for all the samples. Each values were shown as mean ± SD (N = 5), and bars with different letters were significantly different ( P
    Figure Legend Snippet: The expression of eight miRNAs detected by SYBR Green real-time PCR in oyster haemocytes after bacteria challenge and heat stress, including cgi-miR-8 (A), cgi-miR-12 (B), cgi-miR-100 (C), cgi-miR-125 (D), cgi-miR-1984 (E), scaffold631_909 (F), scaffold42648_5080 (G) and scaffold1599_5643 (H). 5S gene was used as an internal control to calibrate the cDNA template for all the samples. Each values were shown as mean ± SD (N = 5), and bars with different letters were significantly different ( P

    Techniques Used: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction

    5) Product Images from "Group 1 LEA proteins contribute to the desiccation and freeze tolerance of Artemia franciscana embryos during diapause"

    Article Title: Group 1 LEA proteins contribute to the desiccation and freeze tolerance of Artemia franciscana embryos during diapause

    Journal: Cell Stress & Chaperones

    doi: 10.1007/s12192-014-0518-3

    Knock down of group 1 LEA proteins in cysts by RNAi. a AfrLEA1 and GFP cDNA were amplified and dsRNAs were synthesized using PCR products. Reaction products were resolved in 1.5 % agarose and visualized with SYBR®Safe. Lane 1 GeneRuler 100 bp DNA Ladder (Thermo Scientific), sizes indicated on left in base pairs; 2 AfrLEA1 PCR product; 3 AfrLEA1 dsRNA; 4 GFP PCR product; 5 GFP dsRNA. b RT-qPCR was performed in duplicate with RNA from 25 to 30 cysts to determine AfrLEA1 and α-tubulin transcript copy number. AfrLEA1 copy numbers were normalized against α-tubulin and averaged for 4 and 6 cyst samples obtained from females injected with either GFP or AfrLEA1 dsRNA. AfrLEA1 copy number in cysts from females injected with GFP dsRNA was set at 100. Error bars represent standard deviation. The asterisk indicates that the mean copy number of group 1 LEA protein transcripts is statistically different from the group 1 LEA protein transcript copy number in cysts from females injected with GFP dsRNA ( p
    Figure Legend Snippet: Knock down of group 1 LEA proteins in cysts by RNAi. a AfrLEA1 and GFP cDNA were amplified and dsRNAs were synthesized using PCR products. Reaction products were resolved in 1.5 % agarose and visualized with SYBR®Safe. Lane 1 GeneRuler 100 bp DNA Ladder (Thermo Scientific), sizes indicated on left in base pairs; 2 AfrLEA1 PCR product; 3 AfrLEA1 dsRNA; 4 GFP PCR product; 5 GFP dsRNA. b RT-qPCR was performed in duplicate with RNA from 25 to 30 cysts to determine AfrLEA1 and α-tubulin transcript copy number. AfrLEA1 copy numbers were normalized against α-tubulin and averaged for 4 and 6 cyst samples obtained from females injected with either GFP or AfrLEA1 dsRNA. AfrLEA1 copy number in cysts from females injected with GFP dsRNA was set at 100. Error bars represent standard deviation. The asterisk indicates that the mean copy number of group 1 LEA protein transcripts is statistically different from the group 1 LEA protein transcript copy number in cysts from females injected with GFP dsRNA ( p

    Techniques Used: Amplification, Synthesized, Polymerase Chain Reaction, Quantitative RT-PCR, Injection, Standard Deviation

    6) Product Images from "Borrelia burgdorferi Uniquely Regulates Its Motility Genes and Has an Intricate Flagellar Hook-Basal Body Structure ▿"

    Article Title: Borrelia burgdorferi Uniquely Regulates Its Motility Genes and Has an Intricate Flagellar Hook-Basal Body Structure ▿

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.01421-07

    qRT-PCR. Reverse transcription with random hexamer primers generated cDNA using total RNA from the wild type (WT) and SC-E1. Signals from qRT-PCR using specific primers for flaA or flaB were quantified with SYBR green fluorescent dye. The data from one
    Figure Legend Snippet: qRT-PCR. Reverse transcription with random hexamer primers generated cDNA using total RNA from the wild type (WT) and SC-E1. Signals from qRT-PCR using specific primers for flaA or flaB were quantified with SYBR green fluorescent dye. The data from one

    Techniques Used: Quantitative RT-PCR, Random Hexamer Labeling, Generated, SYBR Green Assay

    7) Product Images from "Loop-mediated isothermal amplification for the detection of goose circovirus"

    Article Title: Loop-mediated isothermal amplification for the detection of goose circovirus

    Journal: Virology Journal

    doi: 10.1186/1743-422X-9-110

    Specificity of LAMP (A), (B) and real-time PCR for GCV detection. Panel ( A ) shows green fluorescence after addition of SYBR Green to each reaction mixture under UV light, panel ( B ) electrophoresis of LAMP products in 2% agarose gel stained with GelRed™ DNA staining solution while panel ( C ) real-time PCR specificity plot. A - reference DNA of goose circovirus strain P_1_03, B - DNA of goose hemorrhagic polyomavirus (GHPV) strain 2003, C - Muscovy duck parvovirus strain FM, D - goose parvovirus strain B38, E - Fowl adenovirus type-1 strain CELO.
    Figure Legend Snippet: Specificity of LAMP (A), (B) and real-time PCR for GCV detection. Panel ( A ) shows green fluorescence after addition of SYBR Green to each reaction mixture under UV light, panel ( B ) electrophoresis of LAMP products in 2% agarose gel stained with GelRed™ DNA staining solution while panel ( C ) real-time PCR specificity plot. A - reference DNA of goose circovirus strain P_1_03, B - DNA of goose hemorrhagic polyomavirus (GHPV) strain 2003, C - Muscovy duck parvovirus strain FM, D - goose parvovirus strain B38, E - Fowl adenovirus type-1 strain CELO.

    Techniques Used: Real-time Polymerase Chain Reaction, Fluorescence, SYBR Green Assay, Electrophoresis, Agarose Gel Electrophoresis, Staining

    Sensitivity of LAMP (A), (B) and real-time PCR for GCV detection. Panel (A) shows green fluorescence after addition of SYBR Green to each reaction mixture under UV light, panel (B) electrophoresis of LAMP products in 2% agarose gel stained with GelRed™ DNA staining solution while panel (C) real-time PCR sensitivity plot.
    Figure Legend Snippet: Sensitivity of LAMP (A), (B) and real-time PCR for GCV detection. Panel (A) shows green fluorescence after addition of SYBR Green to each reaction mixture under UV light, panel (B) electrophoresis of LAMP products in 2% agarose gel stained with GelRed™ DNA staining solution while panel (C) real-time PCR sensitivity plot.

    Techniques Used: Real-time Polymerase Chain Reaction, Fluorescence, SYBR Green Assay, Electrophoresis, Agarose Gel Electrophoresis, Staining

    8) Product Images from "Development of a SYBR Green I based real-time RT-PCR assay for detection and quantification of bovine coronavirus"

    Article Title: Development of a SYBR Green I based real-time RT-PCR assay for detection and quantification of bovine coronavirus

    Journal: Molecular and Cellular Probes

    doi: 10.1016/j.mcp.2011.03.001

    Sensitivity of the SYBR Green I based real-time PCR. Amplification plots (cycle number versus fluorescence) of (A) serially diluted DNA plasmid standards [copies/reaction] and (B) serially diluted cell-culture grown BCoV reference strain [TCID 50 /ml]. Standard curves generated from the mean cycle threshold ( C T ) values obtained against the (C) diluted DNA plasmid standards (log 10 copy number) and (D) diluted virus strain (log 10 TCID 50 ). The coefficient of determination ( R 2 ) and the equation of the regression curve ( Y ) were calculated.
    Figure Legend Snippet: Sensitivity of the SYBR Green I based real-time PCR. Amplification plots (cycle number versus fluorescence) of (A) serially diluted DNA plasmid standards [copies/reaction] and (B) serially diluted cell-culture grown BCoV reference strain [TCID 50 /ml]. Standard curves generated from the mean cycle threshold ( C T ) values obtained against the (C) diluted DNA plasmid standards (log 10 copy number) and (D) diluted virus strain (log 10 TCID 50 ). The coefficient of determination ( R 2 ) and the equation of the regression curve ( Y ) were calculated.

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction, Amplification, Fluorescence, Plasmid Preparation, Cell Culture, Generated

    9) Product Images from "Roles of Polypyrimidine Tract Binding Proteins in Major Immediate-Early Gene Expression and Viral Replication of Human Cytomegalovirus ▿"

    Article Title: Roles of Polypyrimidine Tract Binding Proteins in Major Immediate-Early Gene Expression and Viral Replication of Human Cytomegalovirus ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.02407-08

    Detection of IE1 gene expression at the RNA level. (A) Diagram of the IE1 gene containing exon 2 (X2), exon 3 (X3), and exon 4 (X4); introns are between the exons. The positions of primers pX2, pX3L, pX3S, pX3X4, and pRev are shown. (B) RT-PCR. Total RNAs from HEp-2-kdPTB (pgfpIE1-transfected) and HEp-2-pLKO (pgfpIE1-transfected) cells (top) or from HEp-2-kdPTB (pgfpIE1cDNA-transfected) and HEp-2-pLKO (pgfpIE1cDNA-transfected) cells (bottom), as described previously, were transcribed using a specific primer, pRev, with an RT kit (Invitrogen). The cDNAs were then amplified using different pairs of primers, as indicated. pgfpIE1 DNA was used as a control (as it produces a product the same size as the cDNAs from pre-mRNA), and the total RNA (treated with RNase-free DNase I) was used as a negative control for the PCR. The bands below single asterisks represent cDNA amplified from pre-mRNA; the bands above double asterisks represent the cDNA amplified from spliced RNA. (C) Real-time RT-PCR. Total RNA (1 μg) isolated from HEp-2-kdPTB (pgfpIE1-transfected) and HEp-2-pLKO (pgfpIE1-transfected) cells, as described previously, were analyzed by one-step real-time RT-PCR using the QuantiTect SYBR Green RT-PCR kit (Qiagen, Valencia, CA). Water was used as a negative control for subtraction of the background. The three curves in each sample represent triplicate experiments.
    Figure Legend Snippet: Detection of IE1 gene expression at the RNA level. (A) Diagram of the IE1 gene containing exon 2 (X2), exon 3 (X3), and exon 4 (X4); introns are between the exons. The positions of primers pX2, pX3L, pX3S, pX3X4, and pRev are shown. (B) RT-PCR. Total RNAs from HEp-2-kdPTB (pgfpIE1-transfected) and HEp-2-pLKO (pgfpIE1-transfected) cells (top) or from HEp-2-kdPTB (pgfpIE1cDNA-transfected) and HEp-2-pLKO (pgfpIE1cDNA-transfected) cells (bottom), as described previously, were transcribed using a specific primer, pRev, with an RT kit (Invitrogen). The cDNAs were then amplified using different pairs of primers, as indicated. pgfpIE1 DNA was used as a control (as it produces a product the same size as the cDNAs from pre-mRNA), and the total RNA (treated with RNase-free DNase I) was used as a negative control for the PCR. The bands below single asterisks represent cDNA amplified from pre-mRNA; the bands above double asterisks represent the cDNA amplified from spliced RNA. (C) Real-time RT-PCR. Total RNA (1 μg) isolated from HEp-2-kdPTB (pgfpIE1-transfected) and HEp-2-pLKO (pgfpIE1-transfected) cells, as described previously, were analyzed by one-step real-time RT-PCR using the QuantiTect SYBR Green RT-PCR kit (Qiagen, Valencia, CA). Water was used as a negative control for subtraction of the background. The three curves in each sample represent triplicate experiments.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Amplification, Negative Control, Polymerase Chain Reaction, Quantitative RT-PCR, Isolation, SYBR Green Assay

    10) Product Images from "Targeting of mTOR catalytic site inhibits multiple steps of the HIV-1 lifecycle and suppresses HIV-1 viremia in humanized mice"

    Article Title: Targeting of mTOR catalytic site inhibits multiple steps of the HIV-1 lifecycle and suppresses HIV-1 viremia in humanized mice

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1511144112

    INK128 inhibits HIV transcription in U1 cells. U1 cells were cultured in the presence of 10 nM PMA and the indicated concentrations of INK128. After 2 d, cells were collected, mRNA isolated, quantified, reverse transcribed and amplified by real time PCR using unspliced HIV cDNA primer pair US.1a/US.2a and housekeeping [beta]-actin primers. For quantification, standard curves for unspliced HIV cDNA and [beta]-actin sequences were generated by performing 10-fold serial dilutions of mRNA isolated from PBLs acutely infected with HIV BaL. PCR amplification was performed using Quantitect SYBR Green PCR Kit in a LightCycler. Negative controls consisted of mixture reactions without the reverse transcription step. Data are means ± SD of two experiments, expressed as relative HIV mRNA expression compared with cultures containing no INK128 after normalization to [beta]-actin levels.
    Figure Legend Snippet: INK128 inhibits HIV transcription in U1 cells. U1 cells were cultured in the presence of 10 nM PMA and the indicated concentrations of INK128. After 2 d, cells were collected, mRNA isolated, quantified, reverse transcribed and amplified by real time PCR using unspliced HIV cDNA primer pair US.1a/US.2a and housekeeping [beta]-actin primers. For quantification, standard curves for unspliced HIV cDNA and [beta]-actin sequences were generated by performing 10-fold serial dilutions of mRNA isolated from PBLs acutely infected with HIV BaL. PCR amplification was performed using Quantitect SYBR Green PCR Kit in a LightCycler. Negative controls consisted of mixture reactions without the reverse transcription step. Data are means ± SD of two experiments, expressed as relative HIV mRNA expression compared with cultures containing no INK128 after normalization to [beta]-actin levels.

    Techniques Used: Cell Culture, Isolation, Amplification, Real-time Polymerase Chain Reaction, Generated, Infection, Polymerase Chain Reaction, SYBR Green Assay, Expressing

    11) Product Images from "Overexpression of Rad51C splice variants in colorectal tumors"

    Article Title: Overexpression of Rad51C splice variants in colorectal tumors

    Journal: Oncotarget

    doi:

    Real time PCR analysis of Rad51C variant expression in LS-174T colorectal tumor cells post 5-azacytidine treatment The LS-174T cells were treated with 5-azacytidine at dose of 5μM for 72 hours. The total RNA was isolated from pre and post 5-azacytidine treated LS-174T colorectal tumor cells and reverse transcribed to cDNA. The cDNA was then used as template for Rad51C variant expression analysis using real time specific primers and SYBR green dye ( Supplementary Table S1 ). The analysis showed 14.3 fold increase in relative expression of RNA for variant 1, 3.4 folds for variant 2 and 4.8 folds for variant 3, and 2.5 folds increase for wild type.
    Figure Legend Snippet: Real time PCR analysis of Rad51C variant expression in LS-174T colorectal tumor cells post 5-azacytidine treatment The LS-174T cells were treated with 5-azacytidine at dose of 5μM for 72 hours. The total RNA was isolated from pre and post 5-azacytidine treated LS-174T colorectal tumor cells and reverse transcribed to cDNA. The cDNA was then used as template for Rad51C variant expression analysis using real time specific primers and SYBR green dye ( Supplementary Table S1 ). The analysis showed 14.3 fold increase in relative expression of RNA for variant 1, 3.4 folds for variant 2 and 4.8 folds for variant 3, and 2.5 folds increase for wild type.

    Techniques Used: Real-time Polymerase Chain Reaction, Variant Assay, Expressing, Isolation, SYBR Green Assay

    Real time PCR analysis of Rad51C variant expression in colorectal tumors and non-tumors The total RNA was isolated from 9 colorectal tumors and non-tumors and reverse transcribed to cDNA. The cDNA was then used as template for Rad51C variant expression analysis using the real time specific primers and SYBR green dye ( Supplementary Table S1 ). On average variant 1 was expressed 4.95 fold higher as compared to matched non-tumors. Wild type, variants 2 and 3 were expressed at lower levels. The expression levels are given as relative RNA levels of each of the three variant in folds normalized by the expression level of the non-tumors.
    Figure Legend Snippet: Real time PCR analysis of Rad51C variant expression in colorectal tumors and non-tumors The total RNA was isolated from 9 colorectal tumors and non-tumors and reverse transcribed to cDNA. The cDNA was then used as template for Rad51C variant expression analysis using the real time specific primers and SYBR green dye ( Supplementary Table S1 ). On average variant 1 was expressed 4.95 fold higher as compared to matched non-tumors. Wild type, variants 2 and 3 were expressed at lower levels. The expression levels are given as relative RNA levels of each of the three variant in folds normalized by the expression level of the non-tumors.

    Techniques Used: Real-time Polymerase Chain Reaction, Variant Assay, Expressing, Isolation, SYBR Green Assay

    12) Product Images from "Association of Clinical Status of Follicular Lymphoma Patients after Autologous Stem Cell Transplant and Quantitative Assessment of Lymphoma in Blood and Bone Marrow as Measured by SYBR Green I Polymerase Chain Reaction"

    Article Title: Association of Clinical Status of Follicular Lymphoma Patients after Autologous Stem Cell Transplant and Quantitative Assessment of Lymphoma in Blood and Bone Marrow as Measured by SYBR Green I Polymerase Chain Reaction

    Journal: The Journal of molecular diagnostics : JMD

    doi: 10.2353/jmoldx.2006.050050

    Assessment of Stem Cell Graft Contamination by SYBR-Green I RQ-PCR
    Figure Legend Snippet: Assessment of Stem Cell Graft Contamination by SYBR-Green I RQ-PCR

    Techniques Used: SYBR Green Assay, Polymerase Chain Reaction

    SYBR Green RQ-PCR: Melt Curve Analysis
    Figure Legend Snippet: SYBR Green RQ-PCR: Melt Curve Analysis

    Techniques Used: SYBR Green Assay, Polymerase Chain Reaction

    13) Product Images from "Detection of Listeria Spp. and Listeria Monocytogenes in Biological Samples by SYBR Green I and TaqMan Probe-based Real-time PCRs"

    Article Title: Detection of Listeria Spp. and Listeria Monocytogenes in Biological Samples by SYBR Green I and TaqMan Probe-based Real-time PCRs

    Journal: Journal of Veterinary Research

    doi: 10.1515/jvetres-2017-0069

    Determination of the sensitivity (a) and linearity (b) of genus-specific SYBR Green I real-time PCR using L23SQ-F/R and Lin23SQ-FR primers for the detection of L. monocytogenes ATCC 13932 strain in the liver samples
    Figure Legend Snippet: Determination of the sensitivity (a) and linearity (b) of genus-specific SYBR Green I real-time PCR using L23SQ-F/R and Lin23SQ-FR primers for the detection of L. monocytogenes ATCC 13932 strain in the liver samples

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction

    Determination of the sensitivity (a) and linearity (b) of species-specific SYBR Green I real-time PCR using hlyA-146-F/R primers for the detection of L. monocytogenes ATCC 13932 strain in the brain samples
    Figure Legend Snippet: Determination of the sensitivity (a) and linearity (b) of species-specific SYBR Green I real-time PCR using hlyA-146-F/R primers for the detection of L. monocytogenes ATCC 13932 strain in the brain samples

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction

    Determination of the sensitivity (a) and linearity (b) of species-specific SYBR Green I real-time PCR using hlyA-177-F/R primers for the detection of L. monocytogenes ATCC 13932 strain in the blood samples
    Figure Legend Snippet: Determination of the sensitivity (a) and linearity (b) of species-specific SYBR Green I real-time PCR using hlyA-177-F/R primers for the detection of L. monocytogenes ATCC 13932 strain in the blood samples

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction

    14) Product Images from "CD1d, a Sentinel Molecule Bridging Innate and Adaptive Immunity, Is Downregulated by the Human Papillomavirus (HPV) E5 Protein: a Possible Mechanism for Immune Evasion by HPV ▿"

    Article Title: CD1d, a Sentinel Molecule Bridging Innate and Adaptive Immunity, Is Downregulated by the Human Papillomavirus (HPV) E5 Protein: a Possible Mechanism for Immune Evasion by HPV ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.01053-10

    CD1d heavy-chain transcription and translation in C33A/CD1d, C33A/CD1d-empty, C33A/CD1d-6E5, and C33A/CD1d-16E5 cells. (A) Transcription of CD1d HC. The mRNA levels of CD1d were analyzed by quantitative RT-PCR using SYBR green methodology. CD1d mRNA levels
    Figure Legend Snippet: CD1d heavy-chain transcription and translation in C33A/CD1d, C33A/CD1d-empty, C33A/CD1d-6E5, and C33A/CD1d-16E5 cells. (A) Transcription of CD1d HC. The mRNA levels of CD1d were analyzed by quantitative RT-PCR using SYBR green methodology. CD1d mRNA levels

    Techniques Used: Quantitative RT-PCR, SYBR Green Assay

    15) Product Images from "Lentiviral Vectors Bearing the Cardiac Promoter of the Na+-Ca2+ Exchanger Report Cardiogenic Differentiation in Stem Cells"

    Article Title: Lentiviral Vectors Bearing the Cardiac Promoter of the Na+-Ca2+ Exchanger Report Cardiogenic Differentiation in Stem Cells

    Journal:

    doi: 10.1038/mt.2008.30

    RT SYBR-Green PCR (quantitative RT-PCR)
    Figure Legend Snippet: RT SYBR-Green PCR (quantitative RT-PCR)

    Techniques Used: SYBR Green Assay, Polymerase Chain Reaction, Quantitative RT-PCR

    16) Product Images from "Retinal Vascular Leakage Occurring in GABA Rho-1 Subunit Deficient Mice"

    Article Title: Retinal Vascular Leakage Occurring in GABA Rho-1 Subunit Deficient Mice

    Journal: Experimental eye research

    doi: 10.1016/j.exer.2010.02.012

    Verification of gene expression modulations using SYBR-Green-I based real-time and quantitative PCR. The house-keeping gene GAPGH was used as normalizing standard. Gene expression fold-changes ( rho-1 +/+ vs rho-1 -/- ) were determined using the ΔΔCt
    Figure Legend Snippet: Verification of gene expression modulations using SYBR-Green-I based real-time and quantitative PCR. The house-keeping gene GAPGH was used as normalizing standard. Gene expression fold-changes ( rho-1 +/+ vs rho-1 -/- ) were determined using the ΔΔCt

    Techniques Used: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction

    17) Product Images from "Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties"

    Article Title: Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0145749

    RT-qPCR of total RNA isolated from superior and inferior spikelets of O . sativa cultivars on various days after anthesis (DAA) for genes identified in the Apical-forward SSH cDNA library. Each bar represents the relative expression (in fold change) of a gene in superior compared with inferior spikelets in Mahalaxmi (A), Upahar (B), OR-1918 (D) and Lalat (E), in inferior spikelets of Upahar compared with inferior spikelets of Mahalaxmi (C) and in inferior spikelets of Lalat compared with inferior spikelets of OR-1918 (F). cDNA was prepared as described in Fig 1 and used as the template for RT-qPCR, which was conducted using QuantiFast SYBR Green PCR Kit (Qiagen) and a Roche LightCycler 480 thermocycler. Each gene, as well as actin as the reference control, was amplified using gene-specific primers ( S2 Table ) designed with Primer Blast. The fold change in expression was calculated by the ΔΔCt method with actin as the reference. Each bar represents the mean ± SD from three independent estimations. The asterisks (*) against each bar represent statistically significant changes in expression (either higher or lower in terms of fold change), at p ≤ 0.05, as determined by the ‘t’ test. The genes examined included 2-oxoglutarate dehydrogenase E1 component ( OGDE1 , LOC_OS04G32020.1), glycogen synthase kinase 3 ( GSK3 ) MsK3 homolog (LOC_OS04G31240.1), hypothetical protein (LOC_OS03G12670.1), sucrose synthase2 ( SUS2 , LOC_OS06G09450.2), AAA+ type ATPase (LOC_OS01G04814.1), and C3HC4 RING finger protein (LOC_OS07G31850.1).
    Figure Legend Snippet: RT-qPCR of total RNA isolated from superior and inferior spikelets of O . sativa cultivars on various days after anthesis (DAA) for genes identified in the Apical-forward SSH cDNA library. Each bar represents the relative expression (in fold change) of a gene in superior compared with inferior spikelets in Mahalaxmi (A), Upahar (B), OR-1918 (D) and Lalat (E), in inferior spikelets of Upahar compared with inferior spikelets of Mahalaxmi (C) and in inferior spikelets of Lalat compared with inferior spikelets of OR-1918 (F). cDNA was prepared as described in Fig 1 and used as the template for RT-qPCR, which was conducted using QuantiFast SYBR Green PCR Kit (Qiagen) and a Roche LightCycler 480 thermocycler. Each gene, as well as actin as the reference control, was amplified using gene-specific primers ( S2 Table ) designed with Primer Blast. The fold change in expression was calculated by the ΔΔCt method with actin as the reference. Each bar represents the mean ± SD from three independent estimations. The asterisks (*) against each bar represent statistically significant changes in expression (either higher or lower in terms of fold change), at p ≤ 0.05, as determined by the ‘t’ test. The genes examined included 2-oxoglutarate dehydrogenase E1 component ( OGDE1 , LOC_OS04G32020.1), glycogen synthase kinase 3 ( GSK3 ) MsK3 homolog (LOC_OS04G31240.1), hypothetical protein (LOC_OS03G12670.1), sucrose synthase2 ( SUS2 , LOC_OS06G09450.2), AAA+ type ATPase (LOC_OS01G04814.1), and C3HC4 RING finger protein (LOC_OS07G31850.1).

    Techniques Used: Quantitative RT-PCR, Isolation, cDNA Library Assay, Expressing, SYBR Green Assay, Polymerase Chain Reaction, Amplification

    RT-PCR of total RNA isolated from superior (A) and inferior (B) spikelets of O. sativa cv. Mahalaxmi panicles on 0, 3 and 6 days after anthesis (DAA or simply D) for analysis of genes overexpressed in the superior compared with inferior spikelets (I) and genes overexpressed in the inferior compared with superior spikelets (II). Total RNA was isolated using TRIzol (Life Technologies) and reverse-transcribed using QuantiTect Reverse Transcription Kit (Qiagen). Each gene was amplified using gene-specific primers designed using Primer Blast. Actin and 18S rRNA were amplified for 30 and 25 cycles, respectively, as positive and loading controls. The other reactions were performed for 30 cycles, and the amplified products were separated on an agarose gel containing ETBR and visualized and photographed using a Gel Doc (Bio-Rad). The primer sequences are provided in S2 Table .
    Figure Legend Snippet: RT-PCR of total RNA isolated from superior (A) and inferior (B) spikelets of O. sativa cv. Mahalaxmi panicles on 0, 3 and 6 days after anthesis (DAA or simply D) for analysis of genes overexpressed in the superior compared with inferior spikelets (I) and genes overexpressed in the inferior compared with superior spikelets (II). Total RNA was isolated using TRIzol (Life Technologies) and reverse-transcribed using QuantiTect Reverse Transcription Kit (Qiagen). Each gene was amplified using gene-specific primers designed using Primer Blast. Actin and 18S rRNA were amplified for 30 and 25 cycles, respectively, as positive and loading controls. The other reactions were performed for 30 cycles, and the amplified products were separated on an agarose gel containing ETBR and visualized and photographed using a Gel Doc (Bio-Rad). The primer sequences are provided in S2 Table .

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Isolation, Amplification, Agarose Gel Electrophoresis

    18) Product Images from "Potent and reversible lentiviral vector restriction in murine induced pluripotent stem cells"

    Article Title: Potent and reversible lentiviral vector restriction in murine induced pluripotent stem cells

    Journal: Retrovirology

    doi: 10.1186/s12977-017-0358-1

    Restriction of HIV-1-based vectors in murine iPSC. a Schematic overview of integrated LV and GV used in this study. Vectors contain two LTR with deleted U3 regions (∆U3, self-inactivating design), flanking an EGFP expression cassette driven by an EFS enhancer/promoter. SD splice donor, Ψ retroviral packaging signal, RRE rev responsive element, SA splice acceptor, cPPT central polypurine tract, wPRE woodchuck hepatitis virus posttranscriptional regulatory element. b Scheme of reprogramming murine fibroblasts into iPSC by retroviral expression of Oct4, Sox2, Klf4 and c-Myc transcription factors. c iPSC transduced with three independently produced viral supernatants of LV and GV at an MOI of 10 and 100 (n = 3). Flow cytometry data of collected EGFP and SSEA1 (pluripotency marker) double positive cells are shown. NTD non-transduced control. One-way ANOVA with Tukey-Kramer post hoc test was used for statistical analyses. ns (not significant) p = 0.258; *** p ≤ 0.001 ( left panel ). LV (n = 6) or GV (n = 4) applied to iPSC at an MOI of 100. Mean vector copy numbers per cell were determined 6–8 days after transduction with SYBR Green-based quantitative real-time PCR, based on EGFP copies, and normalized to endogenous PTBP2 DNA copies. The unpaired t test with Welch’s correction was used for statistical analysis. * p = 0.017 ( right panel ). d LV and GV were applied to different iPSC clones at an MOI of 100. The percentage of EGFP and SSEA1 double positive cells was determined by flow cytometry. One-way ANOVA with Tukey-Kramer post hoc test was used for statistical analyses. #1 ** p = 0.004; #2, #2EX, #3 and #4 *** p ≤ 0.001. e LV and GV from three independently produced supernatants were applied to ESC at an MOI of 10 and 100. Analyses were performed 6–8 days after transduction. The percentage of EGFP and SSEA1 double positive cells was measured by flow cytometry. One-way ANOVA with Tukey-Kramer post hoc test was used for statistical analyses. ns p = 0.660; *** p ≤ 0.001 ( left panel ). Mean vector copy numbers per cell were determined with SYBR Green-based quantitative real-time PCR, based on EGFP copies, and normalized to endogenous PTBP2 DNA copies and a plasmid standard ( right panel )
    Figure Legend Snippet: Restriction of HIV-1-based vectors in murine iPSC. a Schematic overview of integrated LV and GV used in this study. Vectors contain two LTR with deleted U3 regions (∆U3, self-inactivating design), flanking an EGFP expression cassette driven by an EFS enhancer/promoter. SD splice donor, Ψ retroviral packaging signal, RRE rev responsive element, SA splice acceptor, cPPT central polypurine tract, wPRE woodchuck hepatitis virus posttranscriptional regulatory element. b Scheme of reprogramming murine fibroblasts into iPSC by retroviral expression of Oct4, Sox2, Klf4 and c-Myc transcription factors. c iPSC transduced with three independently produced viral supernatants of LV and GV at an MOI of 10 and 100 (n = 3). Flow cytometry data of collected EGFP and SSEA1 (pluripotency marker) double positive cells are shown. NTD non-transduced control. One-way ANOVA with Tukey-Kramer post hoc test was used for statistical analyses. ns (not significant) p = 0.258; *** p ≤ 0.001 ( left panel ). LV (n = 6) or GV (n = 4) applied to iPSC at an MOI of 100. Mean vector copy numbers per cell were determined 6–8 days after transduction with SYBR Green-based quantitative real-time PCR, based on EGFP copies, and normalized to endogenous PTBP2 DNA copies. The unpaired t test with Welch’s correction was used for statistical analysis. * p = 0.017 ( right panel ). d LV and GV were applied to different iPSC clones at an MOI of 100. The percentage of EGFP and SSEA1 double positive cells was determined by flow cytometry. One-way ANOVA with Tukey-Kramer post hoc test was used for statistical analyses. #1 ** p = 0.004; #2, #2EX, #3 and #4 *** p ≤ 0.001. e LV and GV from three independently produced supernatants were applied to ESC at an MOI of 10 and 100. Analyses were performed 6–8 days after transduction. The percentage of EGFP and SSEA1 double positive cells was measured by flow cytometry. One-way ANOVA with Tukey-Kramer post hoc test was used for statistical analyses. ns p = 0.660; *** p ≤ 0.001 ( left panel ). Mean vector copy numbers per cell were determined with SYBR Green-based quantitative real-time PCR, based on EGFP copies, and normalized to endogenous PTBP2 DNA copies and a plasmid standard ( right panel )

    Techniques Used: Expressing, Transduction, Produced, Flow Cytometry, Cytometry, Marker, Plasmid Preparation, SYBR Green Assay, Real-time Polymerase Chain Reaction, Clone Assay

    19) Product Images from "A standard curve based method for relative real time PCR data processing"

    Article Title: A standard curve based method for relative real time PCR data processing

    Journal: BMC Bioinformatics

    doi: 10.1186/1471-2105-6-62

    Optical factors affect the plateau scattering . SYBR Green real time PCR in frosted plates (green) and white plates (blue). Frosted plates cause increased plateau scattering because of inconsistent reflection and refraction (Reproduced from [18], with ABgene ® permission).
    Figure Legend Snippet: Optical factors affect the plateau scattering . SYBR Green real time PCR in frosted plates (green) and white plates (blue). Frosted plates cause increased plateau scattering because of inconsistent reflection and refraction (Reproduced from [18], with ABgene ® permission).

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction

    Effect of staining with SYBR Green 1 on PCR gel . A: Before staining. B: After staining. Before electrophoresis SYBR Green1 was added to marker but not to samples.
    Figure Legend Snippet: Effect of staining with SYBR Green 1 on PCR gel . A: Before staining. B: After staining. Before electrophoresis SYBR Green1 was added to marker but not to samples.

    Techniques Used: Staining, SYBR Green Assay, Polymerase Chain Reaction, Electrophoresis, Marker

    20) Product Images from "Oxygen Glucose Deprivation in Rat Hippocampal Slice Cultures Results in Alterations in Carnitine Homeostasis and Mitochondrial Dysfunction"

    Article Title: Oxygen Glucose Deprivation in Rat Hippocampal Slice Cultures Results in Alterations in Carnitine Homeostasis and Mitochondrial Dysfunction

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0040881

    Oxygen glucose deprivation disrupts carnitine homeostasis in rat hippocampal slice cultures. Rat hippocampal slice cultures were exposed to OGD in the presence or absence of L-carnitine (LCAR, 5 mM, 2 h prior to OGD). Slices were harvested 2 h after OGD and Protein extracts (50 µg) were subjected to Western blot analysis to determine effects on CPT1 (A), CPT2 (B), and CrAT (C) protein levels. A representative blot is shown in the inset of each panel. Two hours after OGD total RNA was also isolated and mRNA levels for CTP1 (D) and CTP2 (E) were determined by SYBR Green real-time RT-PCR analyses. Both protein and mRNA expression was normalized using β-actin. In addition, the effect of OGD, in the presence and absence of LCAR, on free carnitine levels (FC, F) and the acylcarnitine (AC): FC ratio (G) was determined. Data are presented as mean ± S.E from 3 independent experiments using 60 pooled slices per experiment. * = P
    Figure Legend Snippet: Oxygen glucose deprivation disrupts carnitine homeostasis in rat hippocampal slice cultures. Rat hippocampal slice cultures were exposed to OGD in the presence or absence of L-carnitine (LCAR, 5 mM, 2 h prior to OGD). Slices were harvested 2 h after OGD and Protein extracts (50 µg) were subjected to Western blot analysis to determine effects on CPT1 (A), CPT2 (B), and CrAT (C) protein levels. A representative blot is shown in the inset of each panel. Two hours after OGD total RNA was also isolated and mRNA levels for CTP1 (D) and CTP2 (E) were determined by SYBR Green real-time RT-PCR analyses. Both protein and mRNA expression was normalized using β-actin. In addition, the effect of OGD, in the presence and absence of LCAR, on free carnitine levels (FC, F) and the acylcarnitine (AC): FC ratio (G) was determined. Data are presented as mean ± S.E from 3 independent experiments using 60 pooled slices per experiment. * = P

    Techniques Used: Western Blot, Isolation, SYBR Green Assay, Quantitative RT-PCR, Expressing

    21) Product Images from "Development of a SYBR Green I based real-time RT-PCR assay for detection and quantification of bovine coronavirus"

    Article Title: Development of a SYBR Green I based real-time RT-PCR assay for detection and quantification of bovine coronavirus

    Journal: Molecular and Cellular Probes

    doi: 10.1016/j.mcp.2011.03.001

    Sensitivity of the SYBR Green I based real-time PCR. Amplification plots (cycle number versus fluorescence) of (A) serially diluted DNA plasmid standards [copies/reaction] and (B) serially diluted cell-culture grown BCoV reference strain [TCID 50 /ml]. Standard curves generated from the mean cycle threshold ( C T ) values obtained against the (C) diluted DNA plasmid standards (log 10 copy number) and (D) diluted virus strain (log 10 TCID 50 ). The coefficient of determination ( R 2 ) and the equation of the regression curve ( Y ) were calculated.
    Figure Legend Snippet: Sensitivity of the SYBR Green I based real-time PCR. Amplification plots (cycle number versus fluorescence) of (A) serially diluted DNA plasmid standards [copies/reaction] and (B) serially diluted cell-culture grown BCoV reference strain [TCID 50 /ml]. Standard curves generated from the mean cycle threshold ( C T ) values obtained against the (C) diluted DNA plasmid standards (log 10 copy number) and (D) diluted virus strain (log 10 TCID 50 ). The coefficient of determination ( R 2 ) and the equation of the regression curve ( Y ) were calculated.

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction, Amplification, Fluorescence, Plasmid Preparation, Cell Culture, Generated

    22) Product Images from "Effects of CXCR4 Gene Transfer on Cardiac Function After Ischemia-Reperfusion Injury"

    Article Title: Effects of CXCR4 Gene Transfer on Cardiac Function After Ischemia-Reperfusion Injury

    Journal: The American Journal of Pathology

    doi: 10.2353/ajpath.2010.090451

    CXCL12 is upregulated in ischemic myocardium of CXCR4-overexpressed rat. A: CXCL12 protein expression was assessed in hearts injected with either CXCR4 or β-gal and/or control (saline-injected; with IR) by immunofluorescence staining. Frozen sections were fixed and stained with anti-CXCL12 (green) and anti–α actinin (red). Primary Abs visualized with FITC or Texas Red conjugate. Nuclei were stained with DAPI. Images are taken with confocal microscopy. B: Protein lysates were prepared from rats’ hearts one week after CXCR4 gene transfer followed by 30 minutes LAD ligation and 24 hours reperfusion. The ischemic and remote tissues were lysed and analyzed by Western blot. Representative gel of three independent experiments is shown. C: CXCL12 mRNA levels were determined by quantitative real-time PCR (QRT-PCR) using a QuantiTect SYBR Green RT-PCR Kit and using specific primers for CXCL12 and 18S. Primers were designed to generate short amplification products. Densitometric analysis of data from three different experiments is shown. * P
    Figure Legend Snippet: CXCL12 is upregulated in ischemic myocardium of CXCR4-overexpressed rat. A: CXCL12 protein expression was assessed in hearts injected with either CXCR4 or β-gal and/or control (saline-injected; with IR) by immunofluorescence staining. Frozen sections were fixed and stained with anti-CXCL12 (green) and anti–α actinin (red). Primary Abs visualized with FITC or Texas Red conjugate. Nuclei were stained with DAPI. Images are taken with confocal microscopy. B: Protein lysates were prepared from rats’ hearts one week after CXCR4 gene transfer followed by 30 minutes LAD ligation and 24 hours reperfusion. The ischemic and remote tissues were lysed and analyzed by Western blot. Representative gel of three independent experiments is shown. C: CXCL12 mRNA levels were determined by quantitative real-time PCR (QRT-PCR) using a QuantiTect SYBR Green RT-PCR Kit and using specific primers for CXCL12 and 18S. Primers were designed to generate short amplification products. Densitometric analysis of data from three different experiments is shown. * P

    Techniques Used: Expressing, Injection, Immunofluorescence, Staining, Confocal Microscopy, Ligation, Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction, Amplification

    23) Product Images from "Jab1/CSN5 mediates E2F dependent expression of mitotic and apoptotic but not DNA replication targets"

    Article Title: Jab1/CSN5 mediates E2F dependent expression of mitotic and apoptotic but not DNA replication targets

    Journal: Cell Cycle

    doi: 10.4161/cc.10.19.17618

    Figure 3. E2F1 and Jab1/CSN5 co-occupy mitotic and apoptotic target but not DNA replication gene promoters. E2F1 and Jab1 binding to E2F1 target gene promoters was assessed using chromatin IP (ChIP). REF52 cells were cross-linked with formaldehyde, chromatin was isolated and sonicated and immunoprecipitated with anti-E2F1 (A) or anti-Jab1/CSN5 (B) antisera. Precipitated DNA was analyzed by SYBR green real-time PCR using primers and results are displayed as a comparative fold-induced binding from precipitate using no antibody vs. experimental antibody. Specific promoters analyzed are listed below each graph. ChIP results are separated into two parts in section A because of the very different scales for the two classes of genes. (C) Serum deprived quiescent REF52 cells were transfected with E2F1 or vector control. Chromatin was isolated and immunoprecipitated with anti-E2F1 or anti-Jab1 antisera.
    Figure Legend Snippet: Figure 3. E2F1 and Jab1/CSN5 co-occupy mitotic and apoptotic target but not DNA replication gene promoters. E2F1 and Jab1 binding to E2F1 target gene promoters was assessed using chromatin IP (ChIP). REF52 cells were cross-linked with formaldehyde, chromatin was isolated and sonicated and immunoprecipitated with anti-E2F1 (A) or anti-Jab1/CSN5 (B) antisera. Precipitated DNA was analyzed by SYBR green real-time PCR using primers and results are displayed as a comparative fold-induced binding from precipitate using no antibody vs. experimental antibody. Specific promoters analyzed are listed below each graph. ChIP results are separated into two parts in section A because of the very different scales for the two classes of genes. (C) Serum deprived quiescent REF52 cells were transfected with E2F1 or vector control. Chromatin was isolated and immunoprecipitated with anti-E2F1 or anti-Jab1 antisera.

    Techniques Used: Binding Assay, Chromatin Immunoprecipitation, Isolation, Sonication, Immunoprecipitation, SYBR Green Assay, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation

    24) Product Images from "A standard curve based method for relative real time PCR data processing"

    Article Title: A standard curve based method for relative real time PCR data processing

    Journal: BMC Bioinformatics

    doi: 10.1186/1471-2105-6-62

    Optical factors affect the plateau scattering . SYBR Green real time PCR in frosted plates (green) and white plates (blue). Frosted plates cause increased plateau scattering because of inconsistent reflection and refraction (Reproduced from [18], with ABgene ® permission).
    Figure Legend Snippet: Optical factors affect the plateau scattering . SYBR Green real time PCR in frosted plates (green) and white plates (blue). Frosted plates cause increased plateau scattering because of inconsistent reflection and refraction (Reproduced from [18], with ABgene ® permission).

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction

    Effect of staining with SYBR Green 1 on PCR gel . A: Before staining. B: After staining. Before electrophoresis SYBR Green1 was added to marker but not to samples.
    Figure Legend Snippet: Effect of staining with SYBR Green 1 on PCR gel . A: Before staining. B: After staining. Before electrophoresis SYBR Green1 was added to marker but not to samples.

    Techniques Used: Staining, SYBR Green Assay, Polymerase Chain Reaction, Electrophoresis, Marker

    25) Product Images from "Neuroblastoma Derived Secretory Protein (NDSP) Is a Novel Secreted Factor Overexpressed in Neuroblastoma"

    Article Title: Neuroblastoma Derived Secretory Protein (NDSP) Is a Novel Secreted Factor Overexpressed in Neuroblastoma

    Journal: Molecular cancer therapeutics

    doi: 10.1158/1535-7163.MCT-08-1132

    Bioinformatic analysis of NDSP and validation of expression in neuroblastoma. A . NDSP protein sequence aligned to the mouse homologue (A230106N23) and additional human protein (LOC649590). Tan shading denotes sequence identity among all sequences; grey shading denotes sequence identity between human NDSP and the mouse homologue; and yellow shading denotes sequence identity between human NDSP and human LOC649590 protein. B . SYBR Green quantitative PCR was used to compare NDSP mRNA expression in pooled normal tissue cDNA samples to expression in two neuroblastoma tissue samples. ΔΔC T 's were calculated relative to NDSP expression in the small intestine. C . SYBR Green quantitative PCR was used to analyze NDSP mRNA levels in 9 neuroblastoma cell lines compared to the other cancer cell lines that had no detectable NDSP transcript by conventional RT-PCR. D ). Acute lymphocytic leukemia (ALL), alveolar (aRMS) and embryonal (eRMS) rhabdomyosarcoma, Ewing's sarcoma (EW), ependymoma (EP), medulloblastoma (MB), Wilms' tumor (WT), neuroblastoma (NB), and osteosarcoma (OS).
    Figure Legend Snippet: Bioinformatic analysis of NDSP and validation of expression in neuroblastoma. A . NDSP protein sequence aligned to the mouse homologue (A230106N23) and additional human protein (LOC649590). Tan shading denotes sequence identity among all sequences; grey shading denotes sequence identity between human NDSP and the mouse homologue; and yellow shading denotes sequence identity between human NDSP and human LOC649590 protein. B . SYBR Green quantitative PCR was used to compare NDSP mRNA expression in pooled normal tissue cDNA samples to expression in two neuroblastoma tissue samples. ΔΔC T 's were calculated relative to NDSP expression in the small intestine. C . SYBR Green quantitative PCR was used to analyze NDSP mRNA levels in 9 neuroblastoma cell lines compared to the other cancer cell lines that had no detectable NDSP transcript by conventional RT-PCR. D ). Acute lymphocytic leukemia (ALL), alveolar (aRMS) and embryonal (eRMS) rhabdomyosarcoma, Ewing's sarcoma (EW), ependymoma (EP), medulloblastoma (MB), Wilms' tumor (WT), neuroblastoma (NB), and osteosarcoma (OS).

    Techniques Used: Expressing, Sequencing, SYBR Green Assay, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Wilms Tumor Assay

    26) Product Images from "Genome-Wide Profiling of MicroRNAs in Adipose Mesenchymal Stem Cell Differentiation and Mouse Models of Obesity"

    Article Title: Genome-Wide Profiling of MicroRNAs in Adipose Mesenchymal Stem Cell Differentiation and Mouse Models of Obesity

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0021305

    Real-time PCR based profiling of miRNAs being differentially regulated during the course of MSC differentiation to adipocytes. A : RT-PCR based pooling strategy to identify all miRNAs being expressed in any of the samples taken during the differentiation time-course. B : Summary of the expression levels of 583 miRNAs analyzed with the pooling strategy. C : Schematic overview of a typical miRNA PCR amplicon produced by SYBR Green based miScript PCR assays. PCR products are typically around 85 bp long and of these 20 bp resemble the variable miRNA sequence while the remaining 60 bases contain the universal PCR primer binding site introduced during reverse transcription. D–E : Melting curve analysis ( D ) in combination with agarose gel electrophoresis ( E ), Lane 1: miR-21*, lane 2: miR-125a-3p, lane 3: miR-30a PCR-products, “L”: GelPilot 50 bp Ladder revealed the specificity of miRNA PCR products. F : Identification of the most suitable housekeeping gene for the normalization of the miRNA expression data. For this purpose the expression of six small RNAs was analyzed at all time points and in both cell passages. To identify the small RNA showing the lowest variation over the whole sample set the mean values and standard deviations of all time points were calculated for every gene and cell passage. The table shows the standard deviations of these means.
    Figure Legend Snippet: Real-time PCR based profiling of miRNAs being differentially regulated during the course of MSC differentiation to adipocytes. A : RT-PCR based pooling strategy to identify all miRNAs being expressed in any of the samples taken during the differentiation time-course. B : Summary of the expression levels of 583 miRNAs analyzed with the pooling strategy. C : Schematic overview of a typical miRNA PCR amplicon produced by SYBR Green based miScript PCR assays. PCR products are typically around 85 bp long and of these 20 bp resemble the variable miRNA sequence while the remaining 60 bases contain the universal PCR primer binding site introduced during reverse transcription. D–E : Melting curve analysis ( D ) in combination with agarose gel electrophoresis ( E ), Lane 1: miR-21*, lane 2: miR-125a-3p, lane 3: miR-30a PCR-products, “L”: GelPilot 50 bp Ladder revealed the specificity of miRNA PCR products. F : Identification of the most suitable housekeeping gene for the normalization of the miRNA expression data. For this purpose the expression of six small RNAs was analyzed at all time points and in both cell passages. To identify the small RNA showing the lowest variation over the whole sample set the mean values and standard deviations of all time points were calculated for every gene and cell passage. The table shows the standard deviations of these means.

    Techniques Used: Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing, Polymerase Chain Reaction, Amplification, Produced, SYBR Green Assay, Sequencing, Binding Assay, Agarose Gel Electrophoresis

    27) Product Images from "SYBR Green based real-time RT-PCR assay for detection and genotype prediction of bovine noroviruses and assessment of clinical significance in Norway"

    Article Title: SYBR Green based real-time RT-PCR assay for detection and genotype prediction of bovine noroviruses and assessment of clinical significance in Norway

    Journal: Journal of Virological Methods

    doi: 10.1016/j.jviromet.2010.03.028

    Schematic representation of the partial Bo/Newbury2/76/UK genome with the forward (BoNoV72F) and reverse (BoNoV72R2) primer-binding sites (red) for the SYBR Green based real-time RT-PCR assay. Additional primers (CBECU-F and Capsid516R) used for sequencing are depicted in green. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of the article.)
    Figure Legend Snippet: Schematic representation of the partial Bo/Newbury2/76/UK genome with the forward (BoNoV72F) and reverse (BoNoV72R2) primer-binding sites (red) for the SYBR Green based real-time RT-PCR assay. Additional primers (CBECU-F and Capsid516R) used for sequencing are depicted in green. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of the article.)

    Techniques Used: Binding Assay, SYBR Green Assay, Quantitative RT-PCR, Sequencing

    28) Product Images from "Network of Micro RNAs Mediate Translational Repression of Bone Morphogenetic Protein Receptor‐2: Involvement in HIV‐Associated Pulmonary Vascular Remodeling"

    Article Title: Network of Micro RNAs Mediate Translational Repression of Bone Morphogenetic Protein Receptor‐2: Involvement in HIV‐Associated Pulmonary Vascular Remodeling

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    doi: 10.1161/JAHA.117.008472

    Quantitative real‐time ( qRT ) polymerase chain reaction ( PCR ) analyses showing significant upregulation (A) or no change (B) of microRNAs (mi RNA s) that are predicted to target 3′untranslated region of bone morphogenetic protein receptor‐2 in human pulmonary arterial smooth muscle cells ( HPASMC s) exposed to cocaine and/or Tat protein of HIV . HPASMC s were made quiescent with 0.1% fetal bovine serum containing medium for 48 hours followed by a treatment with cocaine at 1 μmol/L and/or HIV Tat at 25 ng/mL for 24, 48, and 72 hours. The total RNA was isolated and cDNA was prepared before proceeding to quantify mi RNA s by SYBR green or Taqman‐based qRT ‐ PCR . * P
    Figure Legend Snippet: Quantitative real‐time ( qRT ) polymerase chain reaction ( PCR ) analyses showing significant upregulation (A) or no change (B) of microRNAs (mi RNA s) that are predicted to target 3′untranslated region of bone morphogenetic protein receptor‐2 in human pulmonary arterial smooth muscle cells ( HPASMC s) exposed to cocaine and/or Tat protein of HIV . HPASMC s were made quiescent with 0.1% fetal bovine serum containing medium for 48 hours followed by a treatment with cocaine at 1 μmol/L and/or HIV Tat at 25 ng/mL for 24, 48, and 72 hours. The total RNA was isolated and cDNA was prepared before proceeding to quantify mi RNA s by SYBR green or Taqman‐based qRT ‐ PCR . * P

    Techniques Used: Polymerase Chain Reaction, Isolation, SYBR Green Assay, Quantitative RT-PCR

    29) Product Images from "Preclinical antitumor activity of ST7612AA1: a new oral thiol-based histone deacetylase (HDAC) inhibitor"

    Article Title: Preclinical antitumor activity of ST7612AA1: a new oral thiol-based histone deacetylase (HDAC) inhibitor

    Journal: Oncotarget

    doi:

    Effect of ST7612AA1 on key molecular targets in colon cancer A) Western Blot analysis for assessing the degree of acetylation of histone H3 and tubulin, and for evaluating the expression levels of various target proteins in HCT-116 tumor xenografts collected 24 hours after the last treatment with 80 mg/10 mL/kg ST7612AA1 (lanes 4-6) once daily, according to the schedule qdx5/wx3w, with respect to vehicle-treated animals (lanes 1-3). Actin is shown as a control for protein loading. Representative blots of tumor samples from 3 animals/group are shown. B) Real-time qPCR analysis of ST7612-induced gene changes in HCT-116 tumor xenografts collected as above described. Data are normalized to cyclophilin A and presented as fold change (average ± s.d.) over the vehicle-treated control mice (n=3 animals/group). Sybr Green-based q-PCR analysis was performed using the primer set shown in Suppl. Table 3 .
    Figure Legend Snippet: Effect of ST7612AA1 on key molecular targets in colon cancer A) Western Blot analysis for assessing the degree of acetylation of histone H3 and tubulin, and for evaluating the expression levels of various target proteins in HCT-116 tumor xenografts collected 24 hours after the last treatment with 80 mg/10 mL/kg ST7612AA1 (lanes 4-6) once daily, according to the schedule qdx5/wx3w, with respect to vehicle-treated animals (lanes 1-3). Actin is shown as a control for protein loading. Representative blots of tumor samples from 3 animals/group are shown. B) Real-time qPCR analysis of ST7612-induced gene changes in HCT-116 tumor xenografts collected as above described. Data are normalized to cyclophilin A and presented as fold change (average ± s.d.) over the vehicle-treated control mice (n=3 animals/group). Sybr Green-based q-PCR analysis was performed using the primer set shown in Suppl. Table 3 .

    Techniques Used: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Mouse Assay, SYBR Green Assay, Polymerase Chain Reaction

    30) Product Images from "Expression of HERV-Fc1, a Human Endogenous Retrovirus, Is Increased in Patients with Active Multiple Sclerosis"

    Article Title: Expression of HERV-Fc1, a Human Endogenous Retrovirus, Is Increased in Patients with Active Multiple Sclerosis

    Journal: Journal of Virology

    doi: 10.1128/JVI.06723-11

    HERV-Fc1 gag DNA copy number calculations in the control group for the two genders. Shown are SYBR green absolute Q-PCR assay analyses of HERV-Fc1-positive healthy controls; HERV-Fc1 gag gene copy numbers are calculated per human genome equivalent (30
    Figure Legend Snippet: HERV-Fc1 gag DNA copy number calculations in the control group for the two genders. Shown are SYBR green absolute Q-PCR assay analyses of HERV-Fc1-positive healthy controls; HERV-Fc1 gag gene copy numbers are calculated per human genome equivalent (30

    Techniques Used: SYBR Green Assay, Polymerase Chain Reaction

    31) Product Images from "Absence of Vsx1 expression in the normal and damaged mouse cornea"

    Article Title: Absence of Vsx1 expression in the normal and damaged mouse cornea

    Journal: Molecular Vision

    doi:

    Representative Vsx1 qRT-PCR amplification plots. Amplification plots of qRT–PCR of wild type mouse retinal ( A ) and corneal ( B ) samples using primers for Vsx1 , Gapdh and Rho shown in Table 1 . x-axis shows qRT–PCR cycle number, y-axis shows SYBR-green fluorescence values.
    Figure Legend Snippet: Representative Vsx1 qRT-PCR amplification plots. Amplification plots of qRT–PCR of wild type mouse retinal ( A ) and corneal ( B ) samples using primers for Vsx1 , Gapdh and Rho shown in Table 1 . x-axis shows qRT–PCR cycle number, y-axis shows SYBR-green fluorescence values.

    Techniques Used: Quantitative RT-PCR, Amplification, SYBR Green Assay, Fluorescence

    32) Product Images from "A Novel Wilms Tumor 1 (WT1) Target Gene Negatively Regulates the WNT Signaling Pathway *"

    Article Title: A Novel Wilms Tumor 1 (WT1) Target Gene Negatively Regulates the WNT Signaling Pathway *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.094334

    WID is a WT1(−KTS) target gene. A , quantitative reverse transcription-PCR analysis of WID. WID transcript level was measured by quantitative reverse transcription-PCR at indicated times after removal of tet in UB27 and UD29 cells. Data are the means ± S.D. of three independent experiments. B , Western blot analysis. Total cell lysates were prepared from UB27 cells (− or + tet) and immunoblotted with α-WT1, α-WID, and α-actin antibodies. C , ChIP analysis. UB27 cells were induced to express WT1(−KTS), and the cross-linked chromatin was immunoprecipitated with α-WT1 or rabbit IgG antibodies, followed by PCR amplification with primers corresponding to the WID enhancer regions E1–E3 ( black boxes ) or to the adjacent regions N1 and N2 ( gray boxes ) located ∼10 kb upstream of the transcriptional start (+ 1 ). Distance between regions is as follows: E1–E2 (420 bp), E2–N1 (1540 bp), N1–E3 (2170 bp), and E3–N2 (1610 bp). Quantitative PCR analysis of ChIP was performed independently using SYBR Green, and the result is presented as the fold increase over IgG ( lower panel ). D , luciferase ( LUC ) reporter assay for WID enhancer regions. NIH3T3 cells were cotransfected with plasmids containing the E1, E2, or E3 regions in either sense ( SE ) or antisense ( AS ) orientations and with either pcDNA3-WT1(−KTS) or empty vector and Renilla luciferase. Data represent the mean ± S.D. from three independent experiments.
    Figure Legend Snippet: WID is a WT1(−KTS) target gene. A , quantitative reverse transcription-PCR analysis of WID. WID transcript level was measured by quantitative reverse transcription-PCR at indicated times after removal of tet in UB27 and UD29 cells. Data are the means ± S.D. of three independent experiments. B , Western blot analysis. Total cell lysates were prepared from UB27 cells (− or + tet) and immunoblotted with α-WT1, α-WID, and α-actin antibodies. C , ChIP analysis. UB27 cells were induced to express WT1(−KTS), and the cross-linked chromatin was immunoprecipitated with α-WT1 or rabbit IgG antibodies, followed by PCR amplification with primers corresponding to the WID enhancer regions E1–E3 ( black boxes ) or to the adjacent regions N1 and N2 ( gray boxes ) located ∼10 kb upstream of the transcriptional start (+ 1 ). Distance between regions is as follows: E1–E2 (420 bp), E2–N1 (1540 bp), N1–E3 (2170 bp), and E3–N2 (1610 bp). Quantitative PCR analysis of ChIP was performed independently using SYBR Green, and the result is presented as the fold increase over IgG ( lower panel ). D , luciferase ( LUC ) reporter assay for WID enhancer regions. NIH3T3 cells were cotransfected with plasmids containing the E1, E2, or E3 regions in either sense ( SE ) or antisense ( AS ) orientations and with either pcDNA3-WT1(−KTS) or empty vector and Renilla luciferase. Data represent the mean ± S.D. from three independent experiments.

    Techniques Used: Polymerase Chain Reaction, Western Blot, Chromatin Immunoprecipitation, Immunoprecipitation, Amplification, Real-time Polymerase Chain Reaction, SYBR Green Assay, Luciferase, Reporter Assay, Plasmid Preparation

    33) Product Images from "Quantitative analysis of cell-type specific gene expression in the green alga Volvox carteri"

    Article Title: Quantitative analysis of cell-type specific gene expression in the green alga Volvox carteri

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-7-321

    Comparison of gene expression of six target genes in gonidia versus somatic cells by quantitative real-time RT-PCR . Amplification curves for A) actA (internal control for the 2 -ΔΔCt method), B) gon167 , C) ssgA , D) rpl37 , E) fer1 , and F) regA . The target-specific fluorescence signal of SYBR Green fluorescence emission (detection range 515–545 nm) is plotted against the number of PCR cycles. Curves of gonidial RT-PCRs are given in red, somatic RT-PCRs in blue. All real-time RT-PCR experiments were carried out in triplicate, and a mean amplification curve was generated for each cell-type. The threshold level is given by a broken, horizontal line. The cycle at which the mean amplification curve of gonidial or somatic real-time RT-PCRs crosses the threshold (C t value) is indicated by a broken, vertical line.
    Figure Legend Snippet: Comparison of gene expression of six target genes in gonidia versus somatic cells by quantitative real-time RT-PCR . Amplification curves for A) actA (internal control for the 2 -ΔΔCt method), B) gon167 , C) ssgA , D) rpl37 , E) fer1 , and F) regA . The target-specific fluorescence signal of SYBR Green fluorescence emission (detection range 515–545 nm) is plotted against the number of PCR cycles. Curves of gonidial RT-PCRs are given in red, somatic RT-PCRs in blue. All real-time RT-PCR experiments were carried out in triplicate, and a mean amplification curve was generated for each cell-type. The threshold level is given by a broken, horizontal line. The cycle at which the mean amplification curve of gonidial or somatic real-time RT-PCRs crosses the threshold (C t value) is indicated by a broken, vertical line.

    Techniques Used: Expressing, Quantitative RT-PCR, Amplification, Fluorescence, SYBR Green Assay, Polymerase Chain Reaction, Generated

    34) Product Images from "Ube2s-stabilized β-catenin protects against myocardial ischemia/reperfusion injury by activating HIF-1α signaling"

    Article Title: Ube2s-stabilized β-catenin protects against myocardial ischemia/reperfusion injury by activating HIF-1α signaling

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.102960

    Ube2s expression is upregulated after MI/R injury. ( A ) qRT-PCR analysis of the mRNA level of Ube2s in the heart from C57BL/6 mice following 12 h, 24 h and 48 h of MI/R injury. Samples from mice receiving sham surgery were used as controls. Each group includes 8 mice. The results were normalized to β-Actin and expressed as relative to sham group. Data are mean ± SD. Data were compared with sham group using one-way ANOVA analysis. **, P
    Figure Legend Snippet: Ube2s expression is upregulated after MI/R injury. ( A ) qRT-PCR analysis of the mRNA level of Ube2s in the heart from C57BL/6 mice following 12 h, 24 h and 48 h of MI/R injury. Samples from mice receiving sham surgery were used as controls. Each group includes 8 mice. The results were normalized to β-Actin and expressed as relative to sham group. Data are mean ± SD. Data were compared with sham group using one-way ANOVA analysis. **, P

    Techniques Used: Expressing, Quantitative RT-PCR, Mouse Assay

    35) Product Images from "Tissue-Specific Methylation of Human Insulin Gene and PCR Assay for Monitoring Beta Cell Death"

    Article Title: Tissue-Specific Methylation of Human Insulin Gene and PCR Assay for Monitoring Beta Cell Death

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0094591

    Primer selection and analytical performance of methylation-specific PCR. A) Schematic illustration of the human INS gene promoter region showing the position of the nine CpG sites. Solid arrows represent the bisulfite-specific primers (BSPs) that amplify both methylated and unmethylated DNA. Dashed arrows represent methylation-specific primers (MSPs) that amplify unmethylated DNA only. B) Unmethylated plasmid was serially diluted after bisulfite conversion and analyzed by qMSP using selected primer sets. Agarose gel electrophoresis of MSP reactions showing the size of the PCR products. C) Graphs of real-time SYBR Green PCR data showing linearity of C q versus log copy number of unmethylated plasmid (averages and standard deviation (SD)) from 5 to 10 6 copies.
    Figure Legend Snippet: Primer selection and analytical performance of methylation-specific PCR. A) Schematic illustration of the human INS gene promoter region showing the position of the nine CpG sites. Solid arrows represent the bisulfite-specific primers (BSPs) that amplify both methylated and unmethylated DNA. Dashed arrows represent methylation-specific primers (MSPs) that amplify unmethylated DNA only. B) Unmethylated plasmid was serially diluted after bisulfite conversion and analyzed by qMSP using selected primer sets. Agarose gel electrophoresis of MSP reactions showing the size of the PCR products. C) Graphs of real-time SYBR Green PCR data showing linearity of C q versus log copy number of unmethylated plasmid (averages and standard deviation (SD)) from 5 to 10 6 copies.

    Techniques Used: Selection, Methylation, Polymerase Chain Reaction, Plasmid Preparation, Agarose Gel Electrophoresis, SYBR Green Assay, Standard Deviation

    36) Product Images from "A standard curve based method for relative real time PCR data processing"

    Article Title: A standard curve based method for relative real time PCR data processing

    Journal: BMC Bioinformatics

    doi: 10.1186/1471-2105-6-62

    Optical factors affect the plateau scattering . SYBR Green real time PCR in frosted plates (green) and white plates (blue). Frosted plates cause increased plateau scattering because of inconsistent reflection and refraction (Reproduced from [18], with ABgene ® permission).
    Figure Legend Snippet: Optical factors affect the plateau scattering . SYBR Green real time PCR in frosted plates (green) and white plates (blue). Frosted plates cause increased plateau scattering because of inconsistent reflection and refraction (Reproduced from [18], with ABgene ® permission).

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction

    Effect of staining with SYBR Green 1 on PCR gel . A: Before staining. B: After staining. Before electrophoresis SYBR Green1 was added to marker but not to samples.
    Figure Legend Snippet: Effect of staining with SYBR Green 1 on PCR gel . A: Before staining. B: After staining. Before electrophoresis SYBR Green1 was added to marker but not to samples.

    Techniques Used: Staining, SYBR Green Assay, Polymerase Chain Reaction, Electrophoresis, Marker

    37) Product Images from "Genetic polymorphism directs IL-6 expression in fibroblasts but not selected other cell types"

    Article Title: Genetic polymorphism directs IL-6 expression in fibroblasts but not selected other cell types

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1520861112

    Elevated unspliced mRNA levels in high-producing fibroblasts supports transcriptional regulation of the differential IL-6 response between synovial fibroblasts. Unspliced ( A and B ) and mature ( C ) IL-6 mRNA expression for two high IL-6–expressing (OA4, OA6) and two low-expressing (RA341, OA502) was determined by qRT-PCR and normalized to the housekeeping gene HPRT. Unspliced primer 1 ( A ) amplifies a region of intron 1. Unspliced primer 2 ( B ) amplifies a region of intron 2. Unspliced mRNA expression averaged 3.5 ± 1.6% of mature mRNA. Primers directed against the IL-6 proximal promoter showed low contamination with genomic DNA, averaging 1.7 ± 2.4% of unspliced mRNA.
    Figure Legend Snippet: Elevated unspliced mRNA levels in high-producing fibroblasts supports transcriptional regulation of the differential IL-6 response between synovial fibroblasts. Unspliced ( A and B ) and mature ( C ) IL-6 mRNA expression for two high IL-6–expressing (OA4, OA6) and two low-expressing (RA341, OA502) was determined by qRT-PCR and normalized to the housekeeping gene HPRT. Unspliced primer 1 ( A ) amplifies a region of intron 1. Unspliced primer 2 ( B ) amplifies a region of intron 2. Unspliced mRNA expression averaged 3.5 ± 1.6% of mature mRNA. Primers directed against the IL-6 proximal promoter showed low contamination with genomic DNA, averaging 1.7 ± 2.4% of unspliced mRNA.

    Techniques Used: Expressing, Quantitative RT-PCR

    38) Product Images from "Knockout Analysis of Arabidopsis Transcription Factors TGA2, TGA5, and TGA6 Reveals Their Redundant and Essential Roles in Systemic Acquired Resistance"

    Article Title: Knockout Analysis of Arabidopsis Transcription Factors TGA2, TGA5, and TGA6 Reveals Their Redundant and Essential Roles in Systemic Acquired Resistance

    Journal: The Plant Cell

    doi: 10.1105/tpc.014894

    PR-1 Expression in Wild-Type, tga6-1 , tga2-1 tga5-1 , and tga6-1 tga2-1 tga5-1 Plants in Response to Treatment with INA. Total RNA was extracted from 20-day-old seedlings grown on MS medium in the presence (+) or absence (−) of 50 μM INA. Relative levels of PR-1 were determined by real-time PCR using SYBR Green I chemistry. Values were normalized to the expression of ACTIN1 and are expressed relative to the level in wild-type plants. This experiment was repeated twice with similar results.
    Figure Legend Snippet: PR-1 Expression in Wild-Type, tga6-1 , tga2-1 tga5-1 , and tga6-1 tga2-1 tga5-1 Plants in Response to Treatment with INA. Total RNA was extracted from 20-day-old seedlings grown on MS medium in the presence (+) or absence (−) of 50 μM INA. Relative levels of PR-1 were determined by real-time PCR using SYBR Green I chemistry. Values were normalized to the expression of ACTIN1 and are expressed relative to the level in wild-type plants. This experiment was repeated twice with similar results.

    Techniques Used: Expressing, Mass Spectrometry, Real-time Polymerase Chain Reaction, SYBR Green Assay

    39) Product Images from "CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4+ T reg cells"

    Article Title: CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4+ T reg cells

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20060772

    ChIP-chip and ChIP-qPCR analysis of FoxP3 bound DNA from CD4 + CD25 hi human T reg cells. Anti-FoxP3 or control rabbit Ig was used to precipitate cross-linked protein–DNA complexes from expanded CD4 + CD25 hi human T reg cells lysate. The cross-linking of the immunoprecipitated material was removed and protease-treated and the DNA was purified and amplified. The resultant material was hybridized to the whole genome using GeneChip Human tiling 1.0R array set to identify the locations of binding sites for FoxP3. Two sets of graphs: FoxP3 IP versus the Ig control and FoxP3 IP versus Input DNA were generated on the hs.NCBIv35 version of the genome essentially following the method described in Cawley et el. (reference 50 ). (a) Signal enrichment graphs of IL-7R locus (chr5:35863179-35918811). Several regions in IL-7R locus are predicted to be positive (chr5:35892564-35892809 promoter) and negative (chr5:35890618-35890846 2K upstream; chr5:35907667-35907852 Intron 4; chr5:35911721-35911888 intron 7 and exon 8). (b) SYBR green qPCR of IL-7R chromosomal regions. FoxP3 IP versus the IgG fold enrichment ratio was determined from duplicate ChIP assay evaluated in duplicate by real time PCR.
    Figure Legend Snippet: ChIP-chip and ChIP-qPCR analysis of FoxP3 bound DNA from CD4 + CD25 hi human T reg cells. Anti-FoxP3 or control rabbit Ig was used to precipitate cross-linked protein–DNA complexes from expanded CD4 + CD25 hi human T reg cells lysate. The cross-linking of the immunoprecipitated material was removed and protease-treated and the DNA was purified and amplified. The resultant material was hybridized to the whole genome using GeneChip Human tiling 1.0R array set to identify the locations of binding sites for FoxP3. Two sets of graphs: FoxP3 IP versus the Ig control and FoxP3 IP versus Input DNA were generated on the hs.NCBIv35 version of the genome essentially following the method described in Cawley et el. (reference 50 ). (a) Signal enrichment graphs of IL-7R locus (chr5:35863179-35918811). Several regions in IL-7R locus are predicted to be positive (chr5:35892564-35892809 promoter) and negative (chr5:35890618-35890846 2K upstream; chr5:35907667-35907852 Intron 4; chr5:35911721-35911888 intron 7 and exon 8). (b) SYBR green qPCR of IL-7R chromosomal regions. FoxP3 IP versus the IgG fold enrichment ratio was determined from duplicate ChIP assay evaluated in duplicate by real time PCR.

    Techniques Used: Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunoprecipitation, Purification, Amplification, Binding Assay, Generated, SYBR Green Assay

    40) Product Images from "Evaluation of Real-time PCR Based on SYBR Green I Fluorescent Dye for Detection of Bacillus Anthracis Strains in Biological Samples"

    Article Title: Evaluation of Real-time PCR Based on SYBR Green I Fluorescent Dye for Detection of Bacillus Anthracis Strains in Biological Samples

    Journal: Journal of Veterinary Research

    doi: 10.2478/jvetres-2018-0075

    Determination of the sensitivity of SYBR Green I real-time PCR with PA-F/R primers for the detection of B. anthracis strain 1/47 in liver (a) and blood (b) samples using Genomic Mini or Genomic Blood Mini Kits for DNA isolation
    Figure Legend Snippet: Determination of the sensitivity of SYBR Green I real-time PCR with PA-F/R primers for the detection of B. anthracis strain 1/47 in liver (a) and blood (b) samples using Genomic Mini or Genomic Blood Mini Kits for DNA isolation

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction, DNA Extraction

    Determination of the sensitivity of SYBR Green I real-time PCR with rpoB-F/R primers for the detection of B. anthracis strain 1/47 in liver (a) and blood (b) samples using a DNeasy Blood and Tissue Kit for DNA isolation
    Figure Legend Snippet: Determination of the sensitivity of SYBR Green I real-time PCR with rpoB-F/R primers for the detection of B. anthracis strain 1/47 in liver (a) and blood (b) samples using a DNeasy Blood and Tissue Kit for DNA isolation

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction, DNA Extraction

    Determination of the sensitivity of SYBR Green I real-time PCR with capC-F/R primers for the detection of B. anthracis strain 1/47 in liver (a) and blood (b) samples using a DNeasy Blood and Tissue Kit for DNA isolation
    Figure Legend Snippet: Determination of the sensitivity of SYBR Green I real-time PCR with capC-F/R primers for the detection of B. anthracis strain 1/47 in liver (a) and blood (b) samples using a DNeasy Blood and Tissue Kit for DNA isolation

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction, DNA Extraction

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    Real-time Polymerase Chain Reaction:

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    Sequencing:

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    Article Snippet: .. 2.5 Real-time PCR conditions The SYBR Green I based real-time PCR assay was performed on an ABI Prism 7500 Sequence Detection System (Applied Biosystems, Foster City, CA, USA) using QuantiTect SYBR Green PCR Kit (Qiagen). .. The assay was carried out in a total volume of 25 μl reaction mixture prepared in triplicates in 96-well optical reaction plates or MicroAmp® optical tubes (Applied Biosystems).

    SYBR Green Assay:

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    Article Title: Fast detection of Noroviruses using a real-time PCR assay and automated sample preparation
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    Article Title: Loop-mediated isothermal amplification for the detection of goose circovirus
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    Article Title: Group 1 LEA proteins contribute to the desiccation and freeze tolerance of Artemia franciscana embryos during diapause
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    Reverse Transcription Polymerase Chain Reaction:

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    Article Snippet: .. QuantiTect™ SYBR® Green RT-PCR kit was purchased from Qiagen, (Qiagen, Hilden, Germany). .. Amplification is performed in a total volume of 20 μl containing 10 μl of kit-supplied QuantiTect™ SYBR® Green RT-PCR Master mix (including HotStar Taq DNA polymerase, reaction buffer, dNTP mix, and SYBR dye), 0.5 μM of each primer, 1 μl RNase-free water, 0.2 μl kit-supplied QuantiTect RT mix (reverse transcriptase), and 6.8 μl of probe.

    Incubation:

    Article Title: Group 1 LEA proteins contribute to the desiccation and freeze tolerance of Artemia franciscana embryos during diapause
    Article Snippet: .. All RNA preparations were also incubated without reverse transcriptase to ensure the absence of genomic DNA. qPCR was conducted with a QuantiTect® SYBR® Green PCR Kit (Qiagen, Mississauga, ON, Canada) in a Rotor-Gene RG-3000 system (Corbett Research, Sydney, NSW, Australia) using 0.5 μl cDNA as template (King et al. ). ..

    Expressing:

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    Polymerase Chain Reaction:

    Article Title: The Identification and Characteristics of Immune-Related MicroRNAs in Haemocytes of Oyster Crassostrea gigas
    Article Snippet: .. The quantitative real-time PCR was carried out in a total volume of 25.0 µL, containing 12.5 µL of 2x QuantiTect SYBR Green PCR Master Mix (QIAGEN, miScript SYBR Green PCR Kit), 2.5 µL of diluted cDNA, 2.5 µL of each primers (10 mmol L−1 ), and 5.0 µL of RNase-free water. .. Eight miRNA fragments were amplified using specific forward primers ( ) and universal reverse primers, and the universal reverse primers and 5S primer ( ) were used to amplify 5S fragment as an internal control to verify the successful reverse transcription and calibrate the DNA template.

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    Article Title: Development of a SYBR Green I based real-time RT-PCR assay for detection and quantification of bovine coronavirus
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    Article Title: Borrelia burgdorferi Uniquely Regulates Its Motility Genes and Has an Intricate Flagellar Hook-Basal Body Structure ▿
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    Article Title: Loop-mediated isothermal amplification for the detection of goose circovirus
    Article Snippet: .. The reactions were set up on in 0.2 ml OptiAmp® optical tubes with caps (Applied Biosystems, Foster City, California, United States) using Quantitect SYBR Green PCR Kit (Qiagen, Hilden, Germany) with application of outer primers F3 and B3 designed for LAMP. .. The reaction conditions were as follows: 25 μL of total mixture, 12.5 μl of 2x QuantiTect SYBR Green PCR Master Mix, 20 pmol of each F3 and B3 primer, 1 μL of DNA template (~25 ng) which was used in LAMP and deionised water.

    Article Title: Group 1 LEA proteins contribute to the desiccation and freeze tolerance of Artemia franciscana embryos during diapause
    Article Snippet: .. All RNA preparations were also incubated without reverse transcriptase to ensure the absence of genomic DNA. qPCR was conducted with a QuantiTect® SYBR® Green PCR Kit (Qiagen, Mississauga, ON, Canada) in a Rotor-Gene RG-3000 system (Corbett Research, Sydney, NSW, Australia) using 0.5 μl cDNA as template (King et al. ). ..

    Article Title: Friend of GATA suppresses the GATA-induced transcription of hepcidin in hepatocytes through a GATA-regulatory element in the HAMP promoter
    Article Snippet: .. Adult human liver total RNA was purchased from Clontech Laboratories, Inc. Quantification of gene expression was performed by a Rotor Gene 3000 Real-time DNA Detection System (Montreal Biotech, Kirkland, QC, Canada) with QuantiTect SYBRGreen I PCR kits (Qiagen), as described previously ( ). ..

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    Qiagen quantitect sybr green pcr kit
    Quantitative real-time polymerase chain reaction analysis of mRNA levels of apoptotic genes in human lung cancer (HepG2) cells treated with zinc oxide nanoparticles (ZnO NPs) at the concentration of 15 μg/mL for 24 hours. Quantitative real-time polymerase chain reaction was performed by <t>QuantiTect</t> <t>SYBR</t> Green <t>PCR</t> kit using an ABI PRISM 7900HT sequence detection system. The β-actin was used as the internal control to normalize the data. ZnO NP-induced alterations in mRNA levels are expressed in relative quantity compared with those for the respective unexposed control cells. ( A ) p53, ( B ) bax, ( C ) bcl-2, and ( D ) caspase-3. Data represented are mean ± standard deviation of three identical experiments made in triplicate. Note: *Statistically significant difference as compared with the controls ( P
    Quantitect Sybr Green Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 5067 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Quantitative real-time polymerase chain reaction analysis of mRNA levels of apoptotic genes in human lung cancer (HepG2) cells treated with zinc oxide nanoparticles (ZnO NPs) at the concentration of 15 μg/mL for 24 hours. Quantitative real-time polymerase chain reaction was performed by QuantiTect SYBR Green PCR kit using an ABI PRISM 7900HT sequence detection system. The β-actin was used as the internal control to normalize the data. ZnO NP-induced alterations in mRNA levels are expressed in relative quantity compared with those for the respective unexposed control cells. ( A ) p53, ( B ) bax, ( C ) bcl-2, and ( D ) caspase-3. Data represented are mean ± standard deviation of three identical experiments made in triplicate. Note: *Statistically significant difference as compared with the controls ( P

    Journal: International Journal of Nanomedicine

    Article Title: Zinc oxide nanoparticles selectively induce apoptosis in human cancer cells through reactive oxygen species

    doi: 10.2147/IJN.S29129

    Figure Lengend Snippet: Quantitative real-time polymerase chain reaction analysis of mRNA levels of apoptotic genes in human lung cancer (HepG2) cells treated with zinc oxide nanoparticles (ZnO NPs) at the concentration of 15 μg/mL for 24 hours. Quantitative real-time polymerase chain reaction was performed by QuantiTect SYBR Green PCR kit using an ABI PRISM 7900HT sequence detection system. The β-actin was used as the internal control to normalize the data. ZnO NP-induced alterations in mRNA levels are expressed in relative quantity compared with those for the respective unexposed control cells. ( A ) p53, ( B ) bax, ( C ) bcl-2, and ( D ) caspase-3. Data represented are mean ± standard deviation of three identical experiments made in triplicate. Note: *Statistically significant difference as compared with the controls ( P

    Article Snippet: Quantitative real-time PCR was performed by QuantiTect SYBR Green PCR kit (Qiagen) using an ABI PRISM 7900HT Sequence Detection System (Applied Biosystems, Foster City, CA).

    Techniques: Real-time Polymerase Chain Reaction, Concentration Assay, SYBR Green Assay, Polymerase Chain Reaction, Sequencing, Standard Deviation

    T m analysis of real-time PCR with SYBR Green as fluorescent dye. T m analysis allows the differentiation of specific PCR product (amplicon) from non-specific amplification products, such as primer-dimers. Samples: 1 to 7, patient samples (stool); 8, negative control (water); 9, positive control (NoV positive stool). Amplified products from NoV template RNA can be reliably distinguished from non-specific products by different melting points. The melting temperature for the positive samples was 79.0 +/- 0.5°C. Non-specific products melt at lower temperatures.

    Journal: BMC Infectious Diseases

    Article Title: Fast detection of Noroviruses using a real-time PCR assay and automated sample preparation

    doi: 10.1186/1471-2334-4-15

    Figure Lengend Snippet: T m analysis of real-time PCR with SYBR Green as fluorescent dye. T m analysis allows the differentiation of specific PCR product (amplicon) from non-specific amplification products, such as primer-dimers. Samples: 1 to 7, patient samples (stool); 8, negative control (water); 9, positive control (NoV positive stool). Amplified products from NoV template RNA can be reliably distinguished from non-specific products by different melting points. The melting temperature for the positive samples was 79.0 +/- 0.5°C. Non-specific products melt at lower temperatures.

    Article Snippet: QuantiTect™ SYBR® Green RT-PCR kit was purchased from Qiagen, (Qiagen, Hilden, Germany).

    Techniques: Real-time Polymerase Chain Reaction, SYBR Green Assay, Polymerase Chain Reaction, Amplification, Negative Control, Positive Control

    The expression of GATA (2, 3, 4, and 6; panels A–D) and FOG (1 and 2; panels E and F) in the human liver and hepatoma cell lines. Total RNA from hepatocytes was isolated using TRizol (Invitrogen Canada). An aliquot of the total RNA (2 μg) was used as a template to synthesize the first-strand cDNA using the Omniscript reverse transcriptase-PCR system (Qiagen), 5 μl of diluted cDNA (1:6) was then used as a template in the RT-PCRs. Quantification of gene expression was performed by a Rotor Gene 3000 Real-time DNA Detection System (Montreal Biotech, Kirkland, QC, Canada) with QuantiTect SYBRGreen I PCR kits (Qiagen). Amplifications were performed in duplicate using at least three different preparations of first-strand cDNAs for each of the three different RNA extractions while adult human liver total RNA was purchased from Clontech Laboratories, Inc. Expression levels for GATA (dark gray bars) and FOG (light gray bars) were normalized to the housekeeping gene β-actin. The results are presented as mean (±S.E.M.) of the relative mRNA expression. A different superscript indicates a statistically significant difference between means ( a–l P

    Journal: Journal of molecular endocrinology

    Article Title: Friend of GATA suppresses the GATA-induced transcription of hepcidin in hepatocytes through a GATA-regulatory element in the HAMP promoter

    doi: 10.1530/JME-11-0060

    Figure Lengend Snippet: The expression of GATA (2, 3, 4, and 6; panels A–D) and FOG (1 and 2; panels E and F) in the human liver and hepatoma cell lines. Total RNA from hepatocytes was isolated using TRizol (Invitrogen Canada). An aliquot of the total RNA (2 μg) was used as a template to synthesize the first-strand cDNA using the Omniscript reverse transcriptase-PCR system (Qiagen), 5 μl of diluted cDNA (1:6) was then used as a template in the RT-PCRs. Quantification of gene expression was performed by a Rotor Gene 3000 Real-time DNA Detection System (Montreal Biotech, Kirkland, QC, Canada) with QuantiTect SYBRGreen I PCR kits (Qiagen). Amplifications were performed in duplicate using at least three different preparations of first-strand cDNAs for each of the three different RNA extractions while adult human liver total RNA was purchased from Clontech Laboratories, Inc. Expression levels for GATA (dark gray bars) and FOG (light gray bars) were normalized to the housekeeping gene β-actin. The results are presented as mean (±S.E.M.) of the relative mRNA expression. A different superscript indicates a statistically significant difference between means ( a–l P

    Article Snippet: Adult human liver total RNA was purchased from Clontech Laboratories, Inc. Quantification of gene expression was performed by a Rotor Gene 3000 Real-time DNA Detection System (Montreal Biotech, Kirkland, QC, Canada) with QuantiTect SYBRGreen I PCR kits (Qiagen), as described previously ( ).

    Techniques: Expressing, Isolation, Polymerase Chain Reaction

    The expression of eight miRNAs detected by SYBR Green real-time PCR in oyster haemocytes after bacteria challenge and heat stress, including cgi-miR-8 (A), cgi-miR-12 (B), cgi-miR-100 (C), cgi-miR-125 (D), cgi-miR-1984 (E), scaffold631_909 (F), scaffold42648_5080 (G) and scaffold1599_5643 (H). 5S gene was used as an internal control to calibrate the cDNA template for all the samples. Each values were shown as mean ± SD (N = 5), and bars with different letters were significantly different ( P

    Journal: PLoS ONE

    Article Title: The Identification and Characteristics of Immune-Related MicroRNAs in Haemocytes of Oyster Crassostrea gigas

    doi: 10.1371/journal.pone.0088397

    Figure Lengend Snippet: The expression of eight miRNAs detected by SYBR Green real-time PCR in oyster haemocytes after bacteria challenge and heat stress, including cgi-miR-8 (A), cgi-miR-12 (B), cgi-miR-100 (C), cgi-miR-125 (D), cgi-miR-1984 (E), scaffold631_909 (F), scaffold42648_5080 (G) and scaffold1599_5643 (H). 5S gene was used as an internal control to calibrate the cDNA template for all the samples. Each values were shown as mean ± SD (N = 5), and bars with different letters were significantly different ( P

    Article Snippet: The quantitative real-time PCR was carried out in a total volume of 25.0 µL, containing 12.5 µL of 2x QuantiTect SYBR Green PCR Master Mix (QIAGEN, miScript SYBR Green PCR Kit), 2.5 µL of diluted cDNA, 2.5 µL of each primers (10 mmol L−1 ), and 5.0 µL of RNase-free water.

    Techniques: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction