complementary dna cdna synthesis  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Thermo Fisher complementary dna cdna synthesis
    ( A ) 28 S and 18 S rRNA bands. Total RNA extraction as visualised in ethidium bromide stained 2% agarose gel. ( B ) Ethidium bromide-stained gel visualisation of RT–PCR products showing a 207 bp band in the positive samples. Lane 1. Molecular Weight Marker (ΦX174 RF <t>DNA</t> Hae III Digest). Lanes 2–4. Positive samples (three melanoma patients). Lanes 5–7. Negative samples (three melanoma patients). Lane 8. Positive control (SK-mel 28 cell line). Lane 9. Negative control (healthy donor). Lane 10. Negative control (sample without <t>cDNA).</t>
    Complementary Dna Cdna Synthesis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 197 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/complementary dna cdna synthesis/product/Thermo Fisher
    Average 96 stars, based on 197 article reviews
    Price from $9.99 to $1999.99
    complementary dna cdna synthesis - by Bioz Stars, 2020-05
    96/100 stars

    Images

    1) Product Images from "Prognostic significance of the sequential detection of circulating melanoma cells by RT-PCR in high-risk melanoma patients receiving adjuvant interferon"

    Article Title: Prognostic significance of the sequential detection of circulating melanoma cells by RT-PCR in high-risk melanoma patients receiving adjuvant interferon

    Journal: British Journal of Cancer

    doi: 10.1038/sj.bjc.6600419

    ( A ) 28 S and 18 S rRNA bands. Total RNA extraction as visualised in ethidium bromide stained 2% agarose gel. ( B ) Ethidium bromide-stained gel visualisation of RT–PCR products showing a 207 bp band in the positive samples. Lane 1. Molecular Weight Marker (ΦX174 RF DNA Hae III Digest). Lanes 2–4. Positive samples (three melanoma patients). Lanes 5–7. Negative samples (three melanoma patients). Lane 8. Positive control (SK-mel 28 cell line). Lane 9. Negative control (healthy donor). Lane 10. Negative control (sample without cDNA).
    Figure Legend Snippet: ( A ) 28 S and 18 S rRNA bands. Total RNA extraction as visualised in ethidium bromide stained 2% agarose gel. ( B ) Ethidium bromide-stained gel visualisation of RT–PCR products showing a 207 bp band in the positive samples. Lane 1. Molecular Weight Marker (ΦX174 RF DNA Hae III Digest). Lanes 2–4. Positive samples (three melanoma patients). Lanes 5–7. Negative samples (three melanoma patients). Lane 8. Positive control (SK-mel 28 cell line). Lane 9. Negative control (healthy donor). Lane 10. Negative control (sample without cDNA).

    Techniques Used: RNA Extraction, Staining, Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Molecular Weight, Marker, Positive Control, Negative Control

    2) Product Images from "Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1 2"

    Article Title: Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1 2

    Journal: Neoplasia (New York, N.Y.)

    doi:

    JK-6L cells show decreased NF-κB activity on bortezomib and verapamil combination. (A) Proteasomal activity of JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 8 hours. The chymotrypsin-like activity was determined using the luminescence-based Proteasome-Glo assay. Mean values and SD were calculated from quadruplicates. Data represent one of three independently experiments. (B) Western blot analysis of total cell lysates from JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 8 hours. Anti-actin antibody staining served as control for an equal loading. One representative immunoblot of two independent experiments is shown. (C) EMSA was performed with nuclear extracts prepared from JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 8 hours. NF-κB DNA-binding activity was analyzed using IRDye end-labeled probes containing specific binding sites. As control, gel was loaded with free-labeled probes without nuclear extract. The intensity of NF-κB DNA binding activity is displayed as arbitrary light units. Data represent one of two independently performed experiments. (D) Myeloma cell lines were treated with 10 nM bortezomib and 70 µ M verapamil or the combination of both inhibitors in the absence or presence of 20 µ M NF-κB inhibitor PS-1145 for 16 hours. Cells were incubated with the vital dye AlamarBlue. The absorbance (OD) was measured using a spectrophotometer. Only relevant significance values were depicted. (E) Quantitative real-time PCR analysis of total RNA isolated from JK-6L treated with 10nMbortezomib and/or 70 µ M for 8 hours. The diagrams showthe relative IκBa mRNA levels of bortezomib-treated JK-6L cells normalized to β-actin mRNA levels, which served as internal control. cDNA from PBS-treated cells served as reference control. The comparative Ct (DDCT) method for relative quantification of gene expression was used. Mean values and SDwere calculated from triplicates. Data represent one of two independently done experiments. Student's t -test for unpaired heteroscedastic samples was used for statistical analysis. # P
    Figure Legend Snippet: JK-6L cells show decreased NF-κB activity on bortezomib and verapamil combination. (A) Proteasomal activity of JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 8 hours. The chymotrypsin-like activity was determined using the luminescence-based Proteasome-Glo assay. Mean values and SD were calculated from quadruplicates. Data represent one of three independently experiments. (B) Western blot analysis of total cell lysates from JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 8 hours. Anti-actin antibody staining served as control for an equal loading. One representative immunoblot of two independent experiments is shown. (C) EMSA was performed with nuclear extracts prepared from JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 8 hours. NF-κB DNA-binding activity was analyzed using IRDye end-labeled probes containing specific binding sites. As control, gel was loaded with free-labeled probes without nuclear extract. The intensity of NF-κB DNA binding activity is displayed as arbitrary light units. Data represent one of two independently performed experiments. (D) Myeloma cell lines were treated with 10 nM bortezomib and 70 µ M verapamil or the combination of both inhibitors in the absence or presence of 20 µ M NF-κB inhibitor PS-1145 for 16 hours. Cells were incubated with the vital dye AlamarBlue. The absorbance (OD) was measured using a spectrophotometer. Only relevant significance values were depicted. (E) Quantitative real-time PCR analysis of total RNA isolated from JK-6L treated with 10nMbortezomib and/or 70 µ M for 8 hours. The diagrams showthe relative IκBa mRNA levels of bortezomib-treated JK-6L cells normalized to β-actin mRNA levels, which served as internal control. cDNA from PBS-treated cells served as reference control. The comparative Ct (DDCT) method for relative quantification of gene expression was used. Mean values and SDwere calculated from triplicates. Data represent one of two independently done experiments. Student's t -test for unpaired heteroscedastic samples was used for statistical analysis. # P

    Techniques Used: Activity Assay, Glo Assay, Western Blot, Staining, Binding Assay, Labeling, Incubation, Spectrophotometry, Real-time Polymerase Chain Reaction, Isolation, Expressing

    3) Product Images from "Context-dependent roles of MDMX (MDM4) and MDM2 in breast cancer proliferation and circulating tumor cells"

    Article Title: Context-dependent roles of MDMX (MDM4) and MDM2 in breast cancer proliferation and circulating tumor cells

    Journal: Breast Cancer Research : BCR

    doi: 10.1186/s13058-018-1094-8

    MDMX knockdown in primary tumors blocks the transcription of CXCR4 and PTGS2 . The 231.mir30.vector-, 231.sh mdm2 -, and 231.sh mdmx -derived primary tumors were lysed and used for total RNA extraction and complementary DNA synthesis. a Microarray analysis revealed selected tumor metastasis-related genes that were either up- or downregulated in 231.sh mdm2 and 231.sh mdmx compared with 231.mir30 vector. Fold changes were gated either > 2 or
    Figure Legend Snippet: MDMX knockdown in primary tumors blocks the transcription of CXCR4 and PTGS2 . The 231.mir30.vector-, 231.sh mdm2 -, and 231.sh mdmx -derived primary tumors were lysed and used for total RNA extraction and complementary DNA synthesis. a Microarray analysis revealed selected tumor metastasis-related genes that were either up- or downregulated in 231.sh mdm2 and 231.sh mdmx compared with 231.mir30 vector. Fold changes were gated either > 2 or

    Techniques Used: Plasmid Preparation, Derivative Assay, RNA Extraction, DNA Synthesis, Microarray

    4) Product Images from "Control of Maize Vegetative and Reproductive Development, Fertility, and rRNAs Silencing by HISTONE DEACETYLASE 108"

    Article Title: Control of Maize Vegetative and Reproductive Development, Fertility, and rRNAs Silencing by HISTONE DEACETYLASE 108

    Journal: Genetics

    doi: 10.1534/genetics.117.300625

    Expression and chromatin analysis of hda108 target genes controlling patterning in maize leaves. (A) Real-Time PCR analysis of LG2 , LG3 , RS2 , and KN1 expression levels in MA and basal part of the 11th leaf. Wild-type, heterozygous, and homozygous mutant plants of BC5S1 and heterozygous and homozygous mutant plants of BC5σ2 segregating progeny were analyzed. Expression of KN1 was tested also in developing tassels of both progenies. Diagrams represent the values of transcript amount standardized to the transcript amount of GAPC2 gene (for details see Materials and Methods ). The analyses were performed using two independent cDNA preparations and three technical replicates. The relative fold changes are calculated comparing them to the arbitrary unit, which is the transcript level in wild-type plants. Bars represent SD. Asterisks indicate statistically significant changes: * P ≤ 0.05, ** P ≤ 0.01; Student’s t -test. (B) Histone modification analysis at selected loci by Real-Time PCR quantification of ChIP DNA performed with α-H4ac and α-H3K9ac antibodies on chromatin extracted from basal part of the 11th leaf of wild-type and homozygous mutant plants. Data are reported as percentage of chromatin input and are average values from two independent ChIP assays and three PCR repetitions for each ChIP assay. SD are reported. Horizontal line indicates the background signal, measured by omitting antibody during ChIP procedure. Asterisks indicate statistically significant changes: * P ≤ 0.05, ** P ≤ 0.01; Student’s t -test.
    Figure Legend Snippet: Expression and chromatin analysis of hda108 target genes controlling patterning in maize leaves. (A) Real-Time PCR analysis of LG2 , LG3 , RS2 , and KN1 expression levels in MA and basal part of the 11th leaf. Wild-type, heterozygous, and homozygous mutant plants of BC5S1 and heterozygous and homozygous mutant plants of BC5σ2 segregating progeny were analyzed. Expression of KN1 was tested also in developing tassels of both progenies. Diagrams represent the values of transcript amount standardized to the transcript amount of GAPC2 gene (for details see Materials and Methods ). The analyses were performed using two independent cDNA preparations and three technical replicates. The relative fold changes are calculated comparing them to the arbitrary unit, which is the transcript level in wild-type plants. Bars represent SD. Asterisks indicate statistically significant changes: * P ≤ 0.05, ** P ≤ 0.01; Student’s t -test. (B) Histone modification analysis at selected loci by Real-Time PCR quantification of ChIP DNA performed with α-H4ac and α-H3K9ac antibodies on chromatin extracted from basal part of the 11th leaf of wild-type and homozygous mutant plants. Data are reported as percentage of chromatin input and are average values from two independent ChIP assays and three PCR repetitions for each ChIP assay. SD are reported. Horizontal line indicates the background signal, measured by omitting antibody during ChIP procedure. Asterisks indicate statistically significant changes: * P ≤ 0.05, ** P ≤ 0.01; Student’s t -test.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Mutagenesis, Modification, Chromatin Immunoprecipitation, Polymerase Chain Reaction

    5) Product Images from "Stem-Loop RT-qPCR as an Efficient Tool for the Detection and Quantification of Small RNAs in Giardia lamblia"

    Article Title: Stem-Loop RT-qPCR as an Efficient Tool for the Detection and Quantification of Small RNAs in Giardia lamblia

    Journal: Genes

    doi: 10.3390/genes7120131

    Amplification of the snoRNA GlsR17 and its derived miRNA (miR2) by PCR and RT-qPCR. Agarose gels (3%) with the PCR amplification products obtained with primers GslR17-Fw and UniLoop ( A ) and miR2-Fw and UniLoop ( B ) using different concentrations of template cDNA: lane 1: 0.025 ng, lane 2: 0.25 ng, lane 3: 2.5 ng, lane 4: 25 ng, lane 5: 250 ng. Lane M: O’RangeRuler TM 10 bp DNA Ladder (Thermo Scientific). PCR conditions were: 95 °C for 30 s, and 40 cycles of 95 °C for 3 s and 60 °C for 30 s; ( C ) melt curves obtained for GlsR17 and miR2 amplification products by real-time PCR were generated using SYBR Green, and the same primers, template concentrations, and conditions as mentioned in A and B.
    Figure Legend Snippet: Amplification of the snoRNA GlsR17 and its derived miRNA (miR2) by PCR and RT-qPCR. Agarose gels (3%) with the PCR amplification products obtained with primers GslR17-Fw and UniLoop ( A ) and miR2-Fw and UniLoop ( B ) using different concentrations of template cDNA: lane 1: 0.025 ng, lane 2: 0.25 ng, lane 3: 2.5 ng, lane 4: 25 ng, lane 5: 250 ng. Lane M: O’RangeRuler TM 10 bp DNA Ladder (Thermo Scientific). PCR conditions were: 95 °C for 30 s, and 40 cycles of 95 °C for 3 s and 60 °C for 30 s; ( C ) melt curves obtained for GlsR17 and miR2 amplification products by real-time PCR were generated using SYBR Green, and the same primers, template concentrations, and conditions as mentioned in A and B.

    Techniques Used: Amplification, Derivative Assay, Polymerase Chain Reaction, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Generated, SYBR Green Assay

    Stem-loop quantitative reverse transcription PCR (RT-qPCR) design to identify small RNAs of Giardia lamblia and schematic representation of the GlsR17 folding. ( A ) The scheme indicates the formation of complementary DNA (cDNA) from stem-loop-small RNA (step 1) and the quantification of the expression by RT-qPCR (step 2); ( B ) predicted structure of the small nucleolar RNA (snoRNA) GlsR17 based on RNAFold; ( C ) structure of the Stem-loop-GlsR17 probe hybridizing to the target small RNA; the cDNA generated by reverse transcription is shown in green color, the Universal ProbeLibrary (UPL) 21 region in blue, and the sequence specific for capturing the GlsR17 and miR2 RNAs (GACTAT) in yellow.
    Figure Legend Snippet: Stem-loop quantitative reverse transcription PCR (RT-qPCR) design to identify small RNAs of Giardia lamblia and schematic representation of the GlsR17 folding. ( A ) The scheme indicates the formation of complementary DNA (cDNA) from stem-loop-small RNA (step 1) and the quantification of the expression by RT-qPCR (step 2); ( B ) predicted structure of the small nucleolar RNA (snoRNA) GlsR17 based on RNAFold; ( C ) structure of the Stem-loop-GlsR17 probe hybridizing to the target small RNA; the cDNA generated by reverse transcription is shown in green color, the Universal ProbeLibrary (UPL) 21 region in blue, and the sequence specific for capturing the GlsR17 and miR2 RNAs (GACTAT) in yellow.

    Techniques Used: Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Generated, Sequencing

    6) Product Images from "Control of Maize Vegetative and Reproductive Development, Fertility, and rRNAs Silencing by HISTONE DEACETYLASE 108"

    Article Title: Control of Maize Vegetative and Reproductive Development, Fertility, and rRNAs Silencing by HISTONE DEACETYLASE 108

    Journal: Genetics

    doi: 10.1534/genetics.117.300625

    Expression and chromatin analysis of hda108 target genes controlling patterning in maize leaves. (A) Real-Time PCR analysis of LG2 , LG3 , RS2 , and KN1 expression levels in MA and basal part of the 11th leaf. Wild-type, heterozygous, and homozygous mutant plants of BC5S1 and heterozygous and homozygous mutant plants of BC5σ2 segregating progeny were analyzed. Expression of KN1 was tested also in developing tassels of both progenies. Diagrams represent the values of transcript amount standardized to the transcript amount of GAPC2 gene (for details see Materials and Methods ). The analyses were performed using two independent cDNA preparations and three technical replicates. The relative fold changes are calculated comparing them to the arbitrary unit, which is the transcript level in wild-type plants. Bars represent SD. Asterisks indicate statistically significant changes: * P ≤ 0.05, ** P ≤ 0.01; Student’s t -test. (B) Histone modification analysis at selected loci by Real-Time PCR quantification of ChIP DNA performed with α-H4ac and α-H3K9ac antibodies on chromatin extracted from basal part of the 11th leaf of wild-type and homozygous mutant plants. Data are reported as percentage of chromatin input and are average values from two independent ChIP assays and three PCR repetitions for each ChIP assay. SD are reported. Horizontal line indicates the background signal, measured by omitting antibody during ChIP procedure. Asterisks indicate statistically significant changes: * P ≤ 0.05, ** P ≤ 0.01; Student’s t -test.
    Figure Legend Snippet: Expression and chromatin analysis of hda108 target genes controlling patterning in maize leaves. (A) Real-Time PCR analysis of LG2 , LG3 , RS2 , and KN1 expression levels in MA and basal part of the 11th leaf. Wild-type, heterozygous, and homozygous mutant plants of BC5S1 and heterozygous and homozygous mutant plants of BC5σ2 segregating progeny were analyzed. Expression of KN1 was tested also in developing tassels of both progenies. Diagrams represent the values of transcript amount standardized to the transcript amount of GAPC2 gene (for details see Materials and Methods ). The analyses were performed using two independent cDNA preparations and three technical replicates. The relative fold changes are calculated comparing them to the arbitrary unit, which is the transcript level in wild-type plants. Bars represent SD. Asterisks indicate statistically significant changes: * P ≤ 0.05, ** P ≤ 0.01; Student’s t -test. (B) Histone modification analysis at selected loci by Real-Time PCR quantification of ChIP DNA performed with α-H4ac and α-H3K9ac antibodies on chromatin extracted from basal part of the 11th leaf of wild-type and homozygous mutant plants. Data are reported as percentage of chromatin input and are average values from two independent ChIP assays and three PCR repetitions for each ChIP assay. SD are reported. Horizontal line indicates the background signal, measured by omitting antibody during ChIP procedure. Asterisks indicate statistically significant changes: * P ≤ 0.05, ** P ≤ 0.01; Student’s t -test.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Mutagenesis, Modification, Chromatin Immunoprecipitation, Polymerase Chain Reaction

    7) Product Images from "Association between TLR-9 polymorphisms and colon cancer susceptibility in Saudi Arabian female patients"

    Article Title: Association between TLR-9 polymorphisms and colon cancer susceptibility in Saudi Arabian female patients

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S106024

    TLR-9 mRNA expression in colon cancer cells and colon cancer tissues. Notes: Total RNA of tissues was extracted from matching normal and colon cancer tissues, reverse-transcribed into cDNA, and then used to measure TLR-9 mRNA expression with specific primers. TLR-9 expression in colon cancer tissues and matching control tissues is shown as mean ± SD. Abbreviations: cDNA, complementary DNA; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
    Figure Legend Snippet: TLR-9 mRNA expression in colon cancer cells and colon cancer tissues. Notes: Total RNA of tissues was extracted from matching normal and colon cancer tissues, reverse-transcribed into cDNA, and then used to measure TLR-9 mRNA expression with specific primers. TLR-9 expression in colon cancer tissues and matching control tissues is shown as mean ± SD. Abbreviations: cDNA, complementary DNA; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

    Techniques Used: Expressing

    8) Product Images from "Context-dependent roles of MDMX (MDM4) and MDM2 in breast cancer proliferation and circulating tumor cells"

    Article Title: Context-dependent roles of MDMX (MDM4) and MDM2 in breast cancer proliferation and circulating tumor cells

    Journal: Breast Cancer Research : BCR

    doi: 10.1186/s13058-018-1094-8

    MDMX knockdown in primary tumors blocks the transcription of CXCR4 and PTGS2 . The 231.mir30.vector-, 231.sh mdm2 -, and 231.sh mdmx -derived primary tumors were lysed and used for total RNA extraction and complementary DNA synthesis. a Microarray analysis revealed selected tumor metastasis-related genes that were either up- or downregulated in 231.sh mdm2 and 231.sh mdmx compared with 231.mir30 vector. Fold changes were gated either > 2 or
    Figure Legend Snippet: MDMX knockdown in primary tumors blocks the transcription of CXCR4 and PTGS2 . The 231.mir30.vector-, 231.sh mdm2 -, and 231.sh mdmx -derived primary tumors were lysed and used for total RNA extraction and complementary DNA synthesis. a Microarray analysis revealed selected tumor metastasis-related genes that were either up- or downregulated in 231.sh mdm2 and 231.sh mdmx compared with 231.mir30 vector. Fold changes were gated either > 2 or

    Techniques Used: Plasmid Preparation, Derivative Assay, RNA Extraction, DNA Synthesis, Microarray

    9) Product Images from "Whole-genome sequencing of human Pegivirus variant from an Egyptian patient co-infected with hepatitis C virus: a case report"

    Article Title: Whole-genome sequencing of human Pegivirus variant from an Egyptian patient co-infected with hepatitis C virus: a case report

    Journal: Virology Journal

    doi: 10.1186/s12985-019-1242-5

    Identification of human pegivirus (HPgV) infection by Revese transcriptase polymerase chain reaction (RT-PCR). Plasma samples before (lane a ) and after (lane b ) viral treatment with daclatasvir and sofosbuvir. Viral RNA was extracted using viral RNA extraction kit followed by synthesis the complementary DNA (cDNA) with a high-capacity cDNA kit (Applied Biosystems). The PCR reaction targeting the 5′ UTR was performed in the thermal cycle, for 40 cycles to amplify a 173 bp fragment. PCR product was loaded on 2% gel and electrophoresed for 20 min at 120 V and 90 A after that the gel was visualized and imaging using photo-documentation system (Biometria). Lane c is a negative control, and lane d contains a DNA marker (50 bp ladder)
    Figure Legend Snippet: Identification of human pegivirus (HPgV) infection by Revese transcriptase polymerase chain reaction (RT-PCR). Plasma samples before (lane a ) and after (lane b ) viral treatment with daclatasvir and sofosbuvir. Viral RNA was extracted using viral RNA extraction kit followed by synthesis the complementary DNA (cDNA) with a high-capacity cDNA kit (Applied Biosystems). The PCR reaction targeting the 5′ UTR was performed in the thermal cycle, for 40 cycles to amplify a 173 bp fragment. PCR product was loaded on 2% gel and electrophoresed for 20 min at 120 V and 90 A after that the gel was visualized and imaging using photo-documentation system (Biometria). Lane c is a negative control, and lane d contains a DNA marker (50 bp ladder)

    Techniques Used: Infection, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, RNA Extraction, Imaging, Negative Control, Marker

    10) Product Images from "Auraptene Attenuates Malignant Properties of Esophageal Stem-Like Cancer Cells"

    Article Title: Auraptene Attenuates Malignant Properties of Esophageal Stem-Like Cancer Cells

    Journal: Technology in Cancer Research & Treatment

    doi: 10.1177/1533034616650119

    RNA Extraction, Complementary DNA Synthesis, and Quantitative Real-Time Polymerase Chain Reaction
    Figure Legend Snippet: RNA Extraction, Complementary DNA Synthesis, and Quantitative Real-Time Polymerase Chain Reaction

    Techniques Used: RNA Extraction, DNA Synthesis, Real-time Polymerase Chain Reaction

    11) Product Images from "Detecting differential allelic expression using high-resolution melting curve analysis: application to the breast cancer susceptibility gene CHEK2"

    Article Title: Detecting differential allelic expression using high-resolution melting curve analysis: application to the breast cancer susceptibility gene CHEK2

    Journal: BMC Medical Genomics

    doi: 10.1186/1755-8794-4-39

    R plot showing the DAE assay results for the 41 heterozygous individuals enrolled in the study . The level of DAE is calculated by dividing the allelic ratio in cDNA by the corresponding ratio in genomic DNA (log cDNA-log gDNA). Statistical significance for DAE is evaluated using Student's t-test. Evidence for DAE is reached when i) the point estimate of the level of DAE (plotted on the horizontal axis) is greater than 20%, ii) the Student's t-test p-value (plotted on the vertical axis) is ≤ 0.05, and iii) the 95% confidence interval of the point estimate (based on 4 replicate assays) does not include 0. Samples above the horizontal line and outside the hatched area reached the statistical threshold for DAE. In our experiment, four samples met all criteria (Samples 2181, 2498, 2500 and 2666).
    Figure Legend Snippet: R plot showing the DAE assay results for the 41 heterozygous individuals enrolled in the study . The level of DAE is calculated by dividing the allelic ratio in cDNA by the corresponding ratio in genomic DNA (log cDNA-log gDNA). Statistical significance for DAE is evaluated using Student's t-test. Evidence for DAE is reached when i) the point estimate of the level of DAE (plotted on the horizontal axis) is greater than 20%, ii) the Student's t-test p-value (plotted on the vertical axis) is ≤ 0.05, and iii) the 95% confidence interval of the point estimate (based on 4 replicate assays) does not include 0. Samples above the horizontal line and outside the hatched area reached the statistical threshold for DAE. In our experiment, four samples met all criteria (Samples 2181, 2498, 2500 and 2666).

    Techniques Used:

    Non-sense mediated mRNA decay causes differential allelic expression in CHEK2*1100delC carriers . Allelic ratio measurements were performed on genomic DNA (gDNA), cDNA derived from LCLs in standard cell culture condition, and cDNA from LCLs treated with puromycin, an NMD inhibiting agent. (A) For a carrier of the mutation, comparison of gDNA and cDNA melting profiles supports the existence of DAE. Puromycin-cDNA profile resembles gDNA, supporting the role of NMD in the DAE observed in this individual. (B) The wild-type sample shows similar profiles in all three situations. HRM profiles were generated with the R script.
    Figure Legend Snippet: Non-sense mediated mRNA decay causes differential allelic expression in CHEK2*1100delC carriers . Allelic ratio measurements were performed on genomic DNA (gDNA), cDNA derived from LCLs in standard cell culture condition, and cDNA from LCLs treated with puromycin, an NMD inhibiting agent. (A) For a carrier of the mutation, comparison of gDNA and cDNA melting profiles supports the existence of DAE. Puromycin-cDNA profile resembles gDNA, supporting the role of NMD in the DAE observed in this individual. (B) The wild-type sample shows similar profiles in all three situations. HRM profiles were generated with the R script.

    Techniques Used: Expressing, Derivative Assay, Cell Culture, Mutagenesis, Generated

    Related Articles

    Incubation:

    Article Title: Prognostic significance of the sequential detection of circulating melanoma cells by RT-PCR in high-risk melanoma patients receiving adjuvant interferon
    Article Snippet: .. Complementary DNA synthesis For the complementary DNA (cDNA) synthesis 1 μg of RNA was incubated for 5 min at 65°C with 3 μg Random Primer oligonucleotides (Gibco-BRL), in a total volume of 13 μl. .. One mM of each deoxynucleotide triphospate (dNTP) (New England Biolabs), 1× First Strand Buffer and 200 U Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) (Gibco BRL), were added to a final volume of 20 μl. cDNA synthesis was performed for 1 h at 37°C, followed by enzyme denaturation at 95°C for 2 min.

    Amplification:

    Article Title: Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1 2
    Article Snippet: .. Complementary DNA (cDNA) synthesis from 1 µg total RNA was performed using the SuperScript II Reverse Transcriptase (Invitrogen, Karlsruhe, Germany) and amplified with Taq polymerase (NEB, Frankfurt/Main, Germany) using specific primers for XBP-1. .. Polymerase chain reaction (PCR) products were separated by electrophoresis on 2% agarose gel and visualized by ethidium bromide staining.

    DNA Synthesis:

    Article Title: Prognostic significance of the sequential detection of circulating melanoma cells by RT-PCR in high-risk melanoma patients receiving adjuvant interferon
    Article Snippet: .. Complementary DNA synthesis For the complementary DNA (cDNA) synthesis 1 μg of RNA was incubated for 5 min at 65°C with 3 μg Random Primer oligonucleotides (Gibco-BRL), in a total volume of 13 μl. .. One mM of each deoxynucleotide triphospate (dNTP) (New England Biolabs), 1× First Strand Buffer and 200 U Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) (Gibco BRL), were added to a final volume of 20 μl. cDNA synthesis was performed for 1 h at 37°C, followed by enzyme denaturation at 95°C for 2 min.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher taqman advanced mirna cdna synthesis kit
    Real-time PCR validation of Cd-dysregulated miRNAs. <t>TaqMan</t> ® Advanced <t>miRNA</t> assays were used to validate selected Cd-dysregulated miRNAs. ( A ) miR-21-5p; ( B ) miR-34a-5p; ( C ) miR-146b-5p; ( D ) miR-224-5p; ( E ) miR-3084a-3p; ( F ) miR-3084c-3p; ( G ) miR-455-3p; ( H ) miR-423-5p. The comparative CT method was used to determine the fold change (±SEM), and an asterisk (*) indicates a statistically significant change in expression in the Cd-treated group versus the saline control ( N = 6 per group, unpaired t -test, p ≤ 0.05).
    Taqman Advanced Mirna Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taqman advanced mirna cdna synthesis kit/product/Thermo Fisher
    Average 99 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    taqman advanced mirna cdna synthesis kit - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    96
    Thermo Fisher first strand complementary dna
    Validation of AS events in different tissues by RT-PCR. Twenty-two multi-exon genes that underwent AS were validated by RT-PCR. RT-PCR analysis was performed on multiple tissues including those used for <t>RNA-seq.</t> eIF4α6 was used as control. Sl, S.lycopersicum cv Heinz1706; Sp, S.pimpinellifolium LA1589. noRT, negative control using reaction mixture without reverse transcriptase added as templates. Genomic, genomic <t>DNA</t> of LA1589 as PCR templates. M, DNA marker 2 K Plus II (Transgen, Beijing). IR, intron retention; AA, alternative acceptor; AD, alternative donor; ES, exon skipping; Others, AS events with more than one of the four basic types. Function description and primer information for the 22 genes can be found in Additional file 8 : Table S5
    First Strand Complementary Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 156 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/first strand complementary dna/product/Thermo Fisher
    Average 96 stars, based on 156 article reviews
    Price from $9.99 to $1999.99
    first strand complementary dna - by Bioz Stars, 2020-05
    96/100 stars
      Buy from Supplier

    99
    Thermo Fisher cdna synthesis
    Tpk1 and Tpk2 are involved in the regulation of virulence-related gene expression. Cells were grown overnight in YPD at 30°C, washed twice with dH 2 O, diluted to 0.1 OD 600 in 10 mL of Spider medium, and incubated for 4 h at 37°C. Total RNA was extracted using TRIzol and treated with the TURBO <t>DNA-</t> free ™ Kit to remove genomic DNA. <t>cDNA</t> synthesis was carried out using 200 ng of RNA with the high-capacity cDNA reverse transcription kit. Transcript expression levels were analyzed by SYBR® Green PCR Master Mix and normalized to the C. tropicalis ACT1 gene using 2 − Δ Δ Ct method. Asterisks indicate statistically significant differences compared with the wild type using Dunnett's multiple comparison test compared to wild type (* P
    Cdna Synthesis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 7772 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cdna synthesis/product/Thermo Fisher
    Average 99 stars, based on 7772 article reviews
    Price from $9.99 to $1999.99
    cdna synthesis - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    Image Search Results


    Real-time PCR validation of Cd-dysregulated miRNAs. TaqMan ® Advanced miRNA assays were used to validate selected Cd-dysregulated miRNAs. ( A ) miR-21-5p; ( B ) miR-34a-5p; ( C ) miR-146b-5p; ( D ) miR-224-5p; ( E ) miR-3084a-3p; ( F ) miR-3084c-3p; ( G ) miR-455-3p; ( H ) miR-423-5p. The comparative CT method was used to determine the fold change (±SEM), and an asterisk (*) indicates a statistically significant change in expression in the Cd-treated group versus the saline control ( N = 6 per group, unpaired t -test, p ≤ 0.05).

    Journal: Toxics

    Article Title: Cadmium Nephrotoxicity Is Associated with Altered MicroRNA Expression in the Rat Renal Cortex

    doi: 10.3390/toxics6010016

    Figure Lengend Snippet: Real-time PCR validation of Cd-dysregulated miRNAs. TaqMan ® Advanced miRNA assays were used to validate selected Cd-dysregulated miRNAs. ( A ) miR-21-5p; ( B ) miR-34a-5p; ( C ) miR-146b-5p; ( D ) miR-224-5p; ( E ) miR-3084a-3p; ( F ) miR-3084c-3p; ( G ) miR-455-3p; ( H ) miR-423-5p. The comparative CT method was used to determine the fold change (±SEM), and an asterisk (*) indicates a statistically significant change in expression in the Cd-treated group versus the saline control ( N = 6 per group, unpaired t -test, p ≤ 0.05).

    Article Snippet: The cDNA template for PCR was prepared using 10 ng of total RNA sample and the TaqMan® Advanced miRNA cDNA Synthesis kit (Thermo Fisher Scientific) following the manufacturer’s recommended protocol.

    Techniques: Real-time Polymerase Chain Reaction, Expressing

    Validation of AS events in different tissues by RT-PCR. Twenty-two multi-exon genes that underwent AS were validated by RT-PCR. RT-PCR analysis was performed on multiple tissues including those used for RNA-seq. eIF4α6 was used as control. Sl, S.lycopersicum cv Heinz1706; Sp, S.pimpinellifolium LA1589. noRT, negative control using reaction mixture without reverse transcriptase added as templates. Genomic, genomic DNA of LA1589 as PCR templates. M, DNA marker 2 K Plus II (Transgen, Beijing). IR, intron retention; AA, alternative acceptor; AD, alternative donor; ES, exon skipping; Others, AS events with more than one of the four basic types. Function description and primer information for the 22 genes can be found in Additional file 8 : Table S5

    Journal: BMC Genomics

    Article Title: Identification of alternative splicing events by RNA sequencing in early growth tomato fruits

    doi: 10.1186/s12864-015-2128-6

    Figure Lengend Snippet: Validation of AS events in different tissues by RT-PCR. Twenty-two multi-exon genes that underwent AS were validated by RT-PCR. RT-PCR analysis was performed on multiple tissues including those used for RNA-seq. eIF4α6 was used as control. Sl, S.lycopersicum cv Heinz1706; Sp, S.pimpinellifolium LA1589. noRT, negative control using reaction mixture without reverse transcriptase added as templates. Genomic, genomic DNA of LA1589 as PCR templates. M, DNA marker 2 K Plus II (Transgen, Beijing). IR, intron retention; AA, alternative acceptor; AD, alternative donor; ES, exon skipping; Others, AS events with more than one of the four basic types. Function description and primer information for the 22 genes can be found in Additional file 8 : Table S5

    Article Snippet: After genomic DNA in these RNA samples was removed by RNase-free DNase according to the manufacturer’s protocol (New England BioLabs, USA), the total RNA (1 μg per sample) was used to synthesize first strand complementary DNA using the First Strand cDNA Synthesis Kit (Thermo Fisher Scientific, USA).

    Techniques: Reverse Transcription Polymerase Chain Reaction, RNA Sequencing Assay, Negative Control, Polymerase Chain Reaction, Marker

    Tpk1 and Tpk2 are involved in the regulation of virulence-related gene expression. Cells were grown overnight in YPD at 30°C, washed twice with dH 2 O, diluted to 0.1 OD 600 in 10 mL of Spider medium, and incubated for 4 h at 37°C. Total RNA was extracted using TRIzol and treated with the TURBO DNA- free ™ Kit to remove genomic DNA. cDNA synthesis was carried out using 200 ng of RNA with the high-capacity cDNA reverse transcription kit. Transcript expression levels were analyzed by SYBR® Green PCR Master Mix and normalized to the C. tropicalis ACT1 gene using 2 − Δ Δ Ct method. Asterisks indicate statistically significant differences compared with the wild type using Dunnett's multiple comparison test compared to wild type (* P

    Journal: Virulence

    Article Title: Protein kinase A governs growth and virulence in Candida tropicalis

    doi: 10.1080/21505594.2017.1414132

    Figure Lengend Snippet: Tpk1 and Tpk2 are involved in the regulation of virulence-related gene expression. Cells were grown overnight in YPD at 30°C, washed twice with dH 2 O, diluted to 0.1 OD 600 in 10 mL of Spider medium, and incubated for 4 h at 37°C. Total RNA was extracted using TRIzol and treated with the TURBO DNA- free ™ Kit to remove genomic DNA. cDNA synthesis was carried out using 200 ng of RNA with the high-capacity cDNA reverse transcription kit. Transcript expression levels were analyzed by SYBR® Green PCR Master Mix and normalized to the C. tropicalis ACT1 gene using 2 − Δ Δ Ct method. Asterisks indicate statistically significant differences compared with the wild type using Dunnett's multiple comparison test compared to wild type (* P

    Article Snippet: Total RNA was then extracted using TRIzol (Ambion, Carlsbad, CA, USA) and treated with TURBO DNA-free ™ Kit (Thermo Fisher Scientific, Vilnius, Lithuania) to remove genomic DNA. cDNA synthesis was carried out using 200 ng of total RNA with the high-capacity cDNA reverse transcription kit (Thermo Fisher Scientific, Vilnius, Lithuania) in a PCR machine (Biometra, Jena, Germany) following the manufacturer's manual.

    Techniques: Expressing, Incubation, SYBR Green Assay, Polymerase Chain Reaction

    Tpk1 and Tpk2 are involved in the regulation of virulence-related gene expression. Cells were grown overnight in YPD at 30°C, washed twice with dH 2 O, diluted to 0.1 OD 600 in 10 mL of Spider medium, and incubated for 4 h at 37°C. Total RNA was extracted using TRIzol and treated with the TURBO DNA- free ™ Kit to remove genomic DNA. cDNA synthesis was carried out using 200 ng of RNA with the high-capacity cDNA reverse transcription kit. Transcript expression levels were analyzed by SYBR® Green PCR Master Mix and normalized to the C. tropicalis ACT1 gene using 2 − Δ Δ Ct method. Asterisks indicate statistically significant differences compared with the wild type using Dunnett's multiple comparison test compared to wild type (* P

    Journal: Virulence

    Article Title: Protein kinase A governs growth and virulence in Candida tropicalis

    doi: 10.1080/21505594.2017.1414132

    Figure Lengend Snippet: Tpk1 and Tpk2 are involved in the regulation of virulence-related gene expression. Cells were grown overnight in YPD at 30°C, washed twice with dH 2 O, diluted to 0.1 OD 600 in 10 mL of Spider medium, and incubated for 4 h at 37°C. Total RNA was extracted using TRIzol and treated with the TURBO DNA- free ™ Kit to remove genomic DNA. cDNA synthesis was carried out using 200 ng of RNA with the high-capacity cDNA reverse transcription kit. Transcript expression levels were analyzed by SYBR® Green PCR Master Mix and normalized to the C. tropicalis ACT1 gene using 2 − Δ Δ Ct method. Asterisks indicate statistically significant differences compared with the wild type using Dunnett's multiple comparison test compared to wild type (* P

    Article Snippet: Total RNA was then extracted using TRIzol (Ambion, Carlsbad, CA, USA) and treated with TURBO DNA- free ™ Kit (Thermo Fisher Scientific, Vilnius, Lithuania) to remove genomic DNA. cDNA synthesis was carried out using 200 ng of total RNA with the high-capacity cDNA reverse transcription kit (Thermo Fisher Scientific, Vilnius, Lithuania) in a PCR machine (Biometra, Jena, Germany) following the manufacturer's manual.

    Techniques: Expressing, Incubation, SYBR Green Assay, Polymerase Chain Reaction