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Qiagen cdna
Confirmation of αN-catenin III genomic structure. Primers spanning the novel exon and exon 7 amplified a single <t>PCR</t> product from human brain <t>cDNA.</t> This 139 base pair product was consistent with the αN-catenin III genomic structure, which
Cdna, supplied by Qiagen, used in various techniques. Bioz Stars score: 95/100, based on 814 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Regulation of a Novel ?N-Catenin Splice Variant in Schizophrenic Smokers"

Article Title: Regulation of a Novel ?N-Catenin Splice Variant in Schizophrenic Smokers

Journal:

doi: 10.1002/ajmg.b.30679

Confirmation of αN-catenin III genomic structure. Primers spanning the novel exon and exon 7 amplified a single PCR product from human brain cDNA. This 139 base pair product was consistent with the αN-catenin III genomic structure, which
Figure Legend Snippet: Confirmation of αN-catenin III genomic structure. Primers spanning the novel exon and exon 7 amplified a single PCR product from human brain cDNA. This 139 base pair product was consistent with the αN-catenin III genomic structure, which

Techniques Used: Amplification, Polymerase Chain Reaction

2) Product Images from "Candida albicans Sun41p, a Putative Glycosidase, Is Involved in Morphogenesis, Cell Wall Biogenesis, and Biofilm Formation ▿"

Article Title: Candida albicans Sun41p, a Putative Glycosidase, Is Involved in Morphogenesis, Cell Wall Biogenesis, and Biofilm Formation ▿

Journal:

doi: 10.1128/EC.00285-07

Differential regulation in sun41 Δ: total RNA of C. albicans wild-type and sun41 Δ mutant cells growing as hyphae in liquid α-MEM were collected and reverse transcribed to cDNA, which was used as a template in a real-time PCR. The
Figure Legend Snippet: Differential regulation in sun41 Δ: total RNA of C. albicans wild-type and sun41 Δ mutant cells growing as hyphae in liquid α-MEM were collected and reverse transcribed to cDNA, which was used as a template in a real-time PCR. The

Techniques Used: Mutagenesis, Real-time Polymerase Chain Reaction

3) Product Images from "Increased cholesterol 7?-hydroxylase expression and size of the bile acid pool in the lactating rat"

Article Title: Increased cholesterol 7?-hydroxylase expression and size of the bile acid pool in the lactating rat

Journal:

doi: 10.1152/ajpgi.00017.2008.

Expression of sterol 27-hydroxylase (Cyp27a1) mRNA during early and midlactation. Real-time PCR was performed in duplicate on cDNA synthesized from liver total RNA from control (open bars) or PP rats (shaded bars) at 4, 10, 16, or 22 h, n = 5–8
Figure Legend Snippet: Expression of sterol 27-hydroxylase (Cyp27a1) mRNA during early and midlactation. Real-time PCR was performed in duplicate on cDNA synthesized from liver total RNA from control (open bars) or PP rats (shaded bars) at 4, 10, 16, or 22 h, n = 5–8

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Synthesized

Expression of sterol 12α-hydroxylase (Cyp8b1) mRNA and protein in control and midlactation rats. A : real-time PCR was performed in duplicate on cDNA synthesized from liver total RNA from control female rats (open bars) and PP day 10 rats (shaded
Figure Legend Snippet: Expression of sterol 12α-hydroxylase (Cyp8b1) mRNA and protein in control and midlactation rats. A : real-time PCR was performed in duplicate on cDNA synthesized from liver total RNA from control female rats (open bars) and PP day 10 rats (shaded

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Synthesized

Expression of cholesterol 7α-hydroxylase (Cyp7a1) mRNA at early and midlactation. Real-time PCR was performed in duplicate on cDNA synthesized from liver total RNA from control female rats (open bars) and PP rats (shaded bars) at 4, 10, 16, and
Figure Legend Snippet: Expression of cholesterol 7α-hydroxylase (Cyp7a1) mRNA at early and midlactation. Real-time PCR was performed in duplicate on cDNA synthesized from liver total RNA from control female rats (open bars) and PP rats (shaded bars) at 4, 10, 16, and

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Synthesized

4) Product Images from "Feasibility of tumor-derived exosome enrichment in the onco-hematology leukemic model of chronic myeloid leukemia"

Article Title: Feasibility of tumor-derived exosome enrichment in the onco-hematology leukemic model of chronic myeloid leukemia

Journal: International Journal of Molecular Medicine

doi: 10.3892/ijmm.2019.4372

Images obtained during the analysis of dPCR chips by Analysis Suite software. In all the figures, the x-axis corresponds to the VIC wavelength channel. Although no VIC-labeled probe was present in the preparation and no signal resulted emitted in VIC wavelength channel, a baseline signal is naturally present in all the samples and correspond to the background of emission. VIC and FAM are the two wavelengths automatically analyzed by Analysis Suite software and are reported in all the graphs generated by this software. (A) The loaded sample was tumor-enriched exosome cDNA from healthy control number 2. Red dots represent the negative micro-reactions. The y-axis corresponds to FAM (BCR-ABL1 probe label) wavelength emission intensity. In the image, a smear of negative micro-reactions in FAM channel is appreciable (arrow). (B) The loaded sample was tumor-enriched exosome cDNA from healthy control number 2. Red dots represent the negative micro-reactions. Y-axis corresponds to FAM (Y4 probe label) wavelength emission intensity. In the image, a smear of negative micro-reactions in FAM channel is appreciable (arrow). (C) The loaded sample was tumor-enriched exosome cDNA from Ph − control number 4. Red and blue dots represent the negative and the Y4 positive micro-reactions, respectively. Y-axis corresponds to FAM (Y4 probe label) wavelength emission intensity. (D) The loaded sample was tumor-enriched exosome cDNA from CML patient number 10. Red and blue dots represent the negative and the BCR-ABL1 positive micro-reactions, respectively. The y-axis corresponds to FAM (BCR-ABL1 probe label) wavelength emission intensity. dPCR, digital PCR; BCR-ABL1, breakpoint cluster region-proto-oncogene 1 tyrosine-protein kinase; CML, chronic myeloid leukemia; Ph − , Philadelphia chromosome-negative.
Figure Legend Snippet: Images obtained during the analysis of dPCR chips by Analysis Suite software. In all the figures, the x-axis corresponds to the VIC wavelength channel. Although no VIC-labeled probe was present in the preparation and no signal resulted emitted in VIC wavelength channel, a baseline signal is naturally present in all the samples and correspond to the background of emission. VIC and FAM are the two wavelengths automatically analyzed by Analysis Suite software and are reported in all the graphs generated by this software. (A) The loaded sample was tumor-enriched exosome cDNA from healthy control number 2. Red dots represent the negative micro-reactions. The y-axis corresponds to FAM (BCR-ABL1 probe label) wavelength emission intensity. In the image, a smear of negative micro-reactions in FAM channel is appreciable (arrow). (B) The loaded sample was tumor-enriched exosome cDNA from healthy control number 2. Red dots represent the negative micro-reactions. Y-axis corresponds to FAM (Y4 probe label) wavelength emission intensity. In the image, a smear of negative micro-reactions in FAM channel is appreciable (arrow). (C) The loaded sample was tumor-enriched exosome cDNA from Ph − control number 4. Red and blue dots represent the negative and the Y4 positive micro-reactions, respectively. Y-axis corresponds to FAM (Y4 probe label) wavelength emission intensity. (D) The loaded sample was tumor-enriched exosome cDNA from CML patient number 10. Red and blue dots represent the negative and the BCR-ABL1 positive micro-reactions, respectively. The y-axis corresponds to FAM (BCR-ABL1 probe label) wavelength emission intensity. dPCR, digital PCR; BCR-ABL1, breakpoint cluster region-proto-oncogene 1 tyrosine-protein kinase; CML, chronic myeloid leukemia; Ph − , Philadelphia chromosome-negative.

Techniques Used: Digital PCR, Software, Labeling, Generated

5) Product Images from "The Phosphorylation of Vinculin on Tyrosine Residues 100 and 1065, Mediated by Src Kinases, Affects Cell Spreading"

Article Title: The Phosphorylation of Vinculin on Tyrosine Residues 100 and 1065, Mediated by Src Kinases, Affects Cell Spreading

Journal:

doi: 10.1091/mbc.E04-03-0264

Effect of wild-type and vinculin mutant proteins on cell spreading. Vinculin null cells were transfected with a cDNA encoding a puromycin resistant gene alone (control) or were cotransfected with cDNAs encoding for wild-type vinculin (wt) or the vinculin
Figure Legend Snippet: Effect of wild-type and vinculin mutant proteins on cell spreading. Vinculin null cells were transfected with a cDNA encoding a puromycin resistant gene alone (control) or were cotransfected with cDNAs encoding for wild-type vinculin (wt) or the vinculin

Techniques Used: Mutagenesis, Transfection

Double phosphorylation mutant protein Y100F/Y1065F is localized to focal adhesion plaques. NIH 3T3 cells were transfected with either a GFP-wild-type vinculin cDNA (wt) or a GFP-Y100F/Y1065F double mutant cDNA. The cells were fixed, permeabilized, and
Figure Legend Snippet: Double phosphorylation mutant protein Y100F/Y1065F is localized to focal adhesion plaques. NIH 3T3 cells were transfected with either a GFP-wild-type vinculin cDNA (wt) or a GFP-Y100F/Y1065F double mutant cDNA. The cells were fixed, permeabilized, and

Techniques Used: Mutagenesis, Transfection

Phosphorylation site-specific antibody recognizing vinculin when phosphorylated on tyrosine residue 1065 reacts with platelet vinculin. (A) COS-7 cells were cotransfected with a constitutively active c-Src (Y529F) cDNA and with cDNAs encoding for wild-type
Figure Legend Snippet: Phosphorylation site-specific antibody recognizing vinculin when phosphorylated on tyrosine residue 1065 reacts with platelet vinculin. (A) COS-7 cells were cotransfected with a constitutively active c-Src (Y529F) cDNA and with cDNAs encoding for wild-type

Techniques Used:

6) Product Images from "TRPV1 activity and substance P release are required for corneal cold nociception"

Article Title: TRPV1 activity and substance P release are required for corneal cold nociception

Journal: Nature Communications

doi: 10.1038/s41467-019-13536-0

Substance P release is required for corneal cold nociception. a The expression of neuropeptides in mouse TRPM8 + trigeminal ganglionic (TG) neurons based on single-cell RNA-seq data. Each dot represents a single TRPM8 + neuron. Full dataset and methods are available in Nguyen et al. 25 . b Single-cell RT-PCR using intron-spanning primers was performed on individual TRPM8-EGFPf sensory neurons that project to the cornea. All TRPM8 + /TRPV1 + neurons express substance P ( Tac1 ), while fewer TRPM8 + / TRPV1 - neurons express Tac1 . Negative control (−): No reverse transcription reaction on RNA sample from whole TG. Positive control (TG): cDNA from whole TG. c Cold treatments (bath temperature drops from 32 to15 °C) promote the release of substance P from dissociated trigeminal neurons, as revealed by ELISA assays. TRPV1 antagonist AMG9810 (AMG, 300 nM, 3 min pre-treatment before cold stimulations) suppressed the release of substance P, compared with the vehicle control (0.0006% DMSO). Cold-associated release of substance P was also suppressed by pre-desensitization of TRPM8 + neurons using TRPM8 agonist cryosim-3 (10 µM). d Both the reflex blinking and eye closing responses to air flow at 13 °C were significantly reduced in Tac1 −/− mice, compared with WT controls ( n = 5 mice/group). e – f NK1 antagonist alleviates cold nociception. The reflex blinking and eye closing responses to cold (air flow at 13 °C) and cryosim-3 (0.1 nmol in 1 μL) were significantly reduced 30 min after intracisternal injection of NK1 antagonist L733,060 hydrochloride (10 µg in 5 µL, n = 5 mice), compared with the vehicle-treated group (0.9% NaCl, n = 5 mice). g Model for neural pathways encoding corneal cold nociception and allodynia. SP: substance P. Data are expressed as mean ± s.e.m. Statistical analysis by two tailed Student’s t -test. * P
Figure Legend Snippet: Substance P release is required for corneal cold nociception. a The expression of neuropeptides in mouse TRPM8 + trigeminal ganglionic (TG) neurons based on single-cell RNA-seq data. Each dot represents a single TRPM8 + neuron. Full dataset and methods are available in Nguyen et al. 25 . b Single-cell RT-PCR using intron-spanning primers was performed on individual TRPM8-EGFPf sensory neurons that project to the cornea. All TRPM8 + /TRPV1 + neurons express substance P ( Tac1 ), while fewer TRPM8 + / TRPV1 - neurons express Tac1 . Negative control (−): No reverse transcription reaction on RNA sample from whole TG. Positive control (TG): cDNA from whole TG. c Cold treatments (bath temperature drops from 32 to15 °C) promote the release of substance P from dissociated trigeminal neurons, as revealed by ELISA assays. TRPV1 antagonist AMG9810 (AMG, 300 nM, 3 min pre-treatment before cold stimulations) suppressed the release of substance P, compared with the vehicle control (0.0006% DMSO). Cold-associated release of substance P was also suppressed by pre-desensitization of TRPM8 + neurons using TRPM8 agonist cryosim-3 (10 µM). d Both the reflex blinking and eye closing responses to air flow at 13 °C were significantly reduced in Tac1 −/− mice, compared with WT controls ( n = 5 mice/group). e – f NK1 antagonist alleviates cold nociception. The reflex blinking and eye closing responses to cold (air flow at 13 °C) and cryosim-3 (0.1 nmol in 1 μL) were significantly reduced 30 min after intracisternal injection of NK1 antagonist L733,060 hydrochloride (10 µg in 5 µL, n = 5 mice), compared with the vehicle-treated group (0.9% NaCl, n = 5 mice). g Model for neural pathways encoding corneal cold nociception and allodynia. SP: substance P. Data are expressed as mean ± s.e.m. Statistical analysis by two tailed Student’s t -test. * P

Techniques Used: Expressing, RNA Sequencing Assay, Reverse Transcription Polymerase Chain Reaction, Negative Control, Positive Control, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Mouse Assay, Injection, Two Tailed Test

7) Product Images from "Characterization of the Cpx Regulon in Escherichia coli Strain MC4100 ▿"

Article Title: Characterization of the Cpx Regulon in Escherichia coli Strain MC4100 ▿

Journal:

doi: 10.1128/JB.00798-08

qPCR analysis of cpxP , degP , ompF , aroK , ompC , and ppiD transcript levels. RNA was isolated from late log (OD 600 of 0.8) cultures of MC4100 and TR10 ( cpxA24 ) and converted to cDNA. The cDNA was subjected to qPCR analysis, and PCR product was quantified
Figure Legend Snippet: qPCR analysis of cpxP , degP , ompF , aroK , ompC , and ppiD transcript levels. RNA was isolated from late log (OD 600 of 0.8) cultures of MC4100 and TR10 ( cpxA24 ) and converted to cDNA. The cDNA was subjected to qPCR analysis, and PCR product was quantified

Techniques Used: Real-time Polymerase Chain Reaction, Isolation, Polymerase Chain Reaction

8) Product Images from "Molecular Dynamics Simulation Reveals Exposed Residues in the Ligand-Binding Domain of the Low-Density Lipoprotein Receptor that Interacts with Vesicular Stomatitis Virus-G Envelope"

Article Title: Molecular Dynamics Simulation Reveals Exposed Residues in the Ligand-Binding Domain of the Low-Density Lipoprotein Receptor that Interacts with Vesicular Stomatitis Virus-G Envelope

Journal: Viruses

doi: 10.3390/v11111063

Transduction of 293T cells with LDLReGFP supernatant including different substitution or deletions at the 3’ of LDLRcDNA. The packaging of the construct harboring eGFP gene only or 5’-LDLReGFP-3’ cassette with different substitutions or deletion was done as described in the materials and methods section, thereafter we used the same supernatant amount to transduce the 293T cells. ( A ) We analyzed the expression of GFP using flow cytometry and express the transduction efficiency in % of GFP. ( B ) After isolation of RNA from the supernatant and synthesis of the first-strand cDNA, as described in the materials and methods section, the GFP fragment was showed using PCR.
Figure Legend Snippet: Transduction of 293T cells with LDLReGFP supernatant including different substitution or deletions at the 3’ of LDLRcDNA. The packaging of the construct harboring eGFP gene only or 5’-LDLReGFP-3’ cassette with different substitutions or deletion was done as described in the materials and methods section, thereafter we used the same supernatant amount to transduce the 293T cells. ( A ) We analyzed the expression of GFP using flow cytometry and express the transduction efficiency in % of GFP. ( B ) After isolation of RNA from the supernatant and synthesis of the first-strand cDNA, as described in the materials and methods section, the GFP fragment was showed using PCR.

Techniques Used: Transduction, Construct, Expressing, Flow Cytometry, Cytometry, Isolation, Polymerase Chain Reaction

9) Product Images from "FGFR4 and its novel splice form in myogenic cells: interplay of glycosylation and tyrosine phosphorylation"

Article Title: FGFR4 and its novel splice form in myogenic cells: interplay of glycosylation and tyrosine phosphorylation

Journal:

doi: 10.1002/jcp.21365

Schematic representations of: (A) Wild type and (−16) splice form of mouse FGFR4 cDNA; the positions of PCR primers used to identify different portions of the corresponding RNA transcripts are indicated with colored arrows and product sizes are
Figure Legend Snippet: Schematic representations of: (A) Wild type and (−16) splice form of mouse FGFR4 cDNA; the positions of PCR primers used to identify different portions of the corresponding RNA transcripts are indicated with colored arrows and product sizes are

Techniques Used: Polymerase Chain Reaction

10) Product Images from "Suppression of topoisomerase IIα expression and function in human cells decreases chromosomal radiosensitivity"

Article Title: Suppression of topoisomerase IIα expression and function in human cells decreases chromosomal radiosensitivity

Journal: Mutation research

doi: 10.1016/j.mrfmmm.2009.01.003

Western blots of topoisomerase IIα and β-actin in hTERT-RPE cells incubated for 12 h with 1 nM and 0.25 nM siRNA against topo IIα or 1 nM scrambled (Scr) siRNA (A). (B) RT-PCR products (reactions performed in triplicate) from cDNA extracted from control hTERT-RPE cells, and cells treated with siRNA against topo IIα (1 nM, 12 h). Negative control represents an RT-PCR reaction without cDNA and M stand for marker. (C) Immunocytochemistry with hTERT-RPE cells incubated with or without 1 nM siRNA against topo IIα for 12 h using primary anti-topo IIα antibody, and FITC-labelled secondary antibody (right panels). Nuclei (left panels) are stained with DAPI (magnification 63×).
Figure Legend Snippet: Western blots of topoisomerase IIα and β-actin in hTERT-RPE cells incubated for 12 h with 1 nM and 0.25 nM siRNA against topo IIα or 1 nM scrambled (Scr) siRNA (A). (B) RT-PCR products (reactions performed in triplicate) from cDNA extracted from control hTERT-RPE cells, and cells treated with siRNA against topo IIα (1 nM, 12 h). Negative control represents an RT-PCR reaction without cDNA and M stand for marker. (C) Immunocytochemistry with hTERT-RPE cells incubated with or without 1 nM siRNA against topo IIα for 12 h using primary anti-topo IIα antibody, and FITC-labelled secondary antibody (right panels). Nuclei (left panels) are stained with DAPI (magnification 63×).

Techniques Used: Western Blot, Incubation, Reverse Transcription Polymerase Chain Reaction, Negative Control, Marker, Immunocytochemistry, Staining

11) Product Images from "Blockade of multiple monoamines receptors reduce insulin secretion from pancreatic β-cells"

Article Title: Blockade of multiple monoamines receptors reduce insulin secretion from pancreatic β-cells

Journal: Scientific Reports

doi: 10.1038/s41598-019-52590-y

mRNA expression of dopamine (D 2 , D 3 , and D 4 ), serotonin (5-HT 2A , 5-HT 2B , 5-HT 2C , and 5-HT 6 ), and histamine (H 1 and H 2 ) receptors in HIT-T15 cells. Total RNA extracted from HIT-T15 cells was reverse-transcribed, and first-strand cDNA was synthesized. Target genes were amplified with a set of specific primers (shown in Table 2 ). For expression analysis of dopamine D 3 and D 4 , all serotonin, and histamine H 2 receptors, two-step PCR was performed using nested primers (shown in Table 2 ). PCR products were separated by electrophoresis using a 2% agarose gel and stained with ethidium bromide. M, 100-bp ladder size marker.
Figure Legend Snippet: mRNA expression of dopamine (D 2 , D 3 , and D 4 ), serotonin (5-HT 2A , 5-HT 2B , 5-HT 2C , and 5-HT 6 ), and histamine (H 1 and H 2 ) receptors in HIT-T15 cells. Total RNA extracted from HIT-T15 cells was reverse-transcribed, and first-strand cDNA was synthesized. Target genes were amplified with a set of specific primers (shown in Table 2 ). For expression analysis of dopamine D 3 and D 4 , all serotonin, and histamine H 2 receptors, two-step PCR was performed using nested primers (shown in Table 2 ). PCR products were separated by electrophoresis using a 2% agarose gel and stained with ethidium bromide. M, 100-bp ladder size marker.

Techniques Used: Expressing, Synthesized, Amplification, Polymerase Chain Reaction, Electrophoresis, Agarose Gel Electrophoresis, Staining, Marker

Related Articles

Digital PCR:

Article Title: Feasibility of tumor-derived exosome enrichment in the onco-hematology leukemic model of chronic myeloid leukemia
Article Snippet: .. A 16 µ l of reaction mixture containing 8 µ l 2X QuantStudio 3D Digital PCR Master Mix (Thermo Fisher Scientific, Inc.), 0.8 µ l 20X TaqMan-MGB-FAM-probe assay for BCR-ABL1 , 5 µ l diluted cDNA and 2.2 µ l nuclease-free water (Qiagen, Inc.). .. The negative control reaction contained no cDNA and was made up to a total of 16 µ l with 7.2 µ l nuclease-free water; a negative control sample was used for each round of thermocycling (as aforementioned).

Amplification:

Article Title: Regulation of a Novel ?N-Catenin Splice Variant in Schizophrenic Smokers
Article Snippet: .. PCR amplification of cDNA was performed using 500 nM of primers and the Quantitect SYBR Green PCR Kit (Qiagen). .. PCR cycling conditions were 50°C for 2 min, 95°C for 15min, and 40 cycles of 94°C for 15 sec, 58°C for 30 sec, and 72°C for 30 sec.

Synthesized:

Article Title: Increased cholesterol 7?-hydroxylase expression and size of the bile acid pool in the lactating rat
Article Snippet: .. Real-time PCR was performed on a Roche LightCycler with a SYBR Green kit by using cDNA synthesized from total RNA isolated with TRIzol Reagent and further purified with the Qiagen RNeasy Mini kit. .. Serial dilutions of cDNA template (0.1, 0.01, 0.001, and 0.001) were amplified, and the cycle number vs. the log of the fluorescence measurement at the threshold was plotted to generate a standard curve for semiquantitation of mRNA expression.

Isolation:

Article Title: Increased cholesterol 7?-hydroxylase expression and size of the bile acid pool in the lactating rat
Article Snippet: .. Real-time PCR was performed on a Roche LightCycler with a SYBR Green kit by using cDNA synthesized from total RNA isolated with TRIzol Reagent and further purified with the Qiagen RNeasy Mini kit. .. Serial dilutions of cDNA template (0.1, 0.01, 0.001, and 0.001) were amplified, and the cycle number vs. the log of the fluorescence measurement at the threshold was plotted to generate a standard curve for semiquantitation of mRNA expression.

SYBR Green Assay:

Article Title: Regulation of a Novel ?N-Catenin Splice Variant in Schizophrenic Smokers
Article Snippet: .. PCR amplification of cDNA was performed using 500 nM of primers and the Quantitect SYBR Green PCR Kit (Qiagen). .. PCR cycling conditions were 50°C for 2 min, 95°C for 15min, and 40 cycles of 94°C for 15 sec, 58°C for 30 sec, and 72°C for 30 sec.

Article Title: Assessing DNA methylation in the developing human intestinal epithelium: potential link to inflammatory bowel disease
Article Snippet: .. Complementary DNA corresponding to 5 ng RNA was used for real-time PCR using QuantiFast SYBR Green PCR Master Mix (Qiagen) on the 7500 Fast real-time PCR system, 7500 software v2.0.6 (Applied Biosciences by Thermo Fisher Scientific, Paisley, UK). .. Relative quantitation was calculated using the ΔΔC t method following normalization against GAPDH.

Article Title: Increased cholesterol 7?-hydroxylase expression and size of the bile acid pool in the lactating rat
Article Snippet: .. Real-time PCR was performed on a Roche LightCycler with a SYBR Green kit by using cDNA synthesized from total RNA isolated with TRIzol Reagent and further purified with the Qiagen RNeasy Mini kit. .. Serial dilutions of cDNA template (0.1, 0.01, 0.001, and 0.001) were amplified, and the cycle number vs. the log of the fluorescence measurement at the threshold was plotted to generate a standard curve for semiquantitation of mRNA expression.

Purification:

Article Title: Increased cholesterol 7?-hydroxylase expression and size of the bile acid pool in the lactating rat
Article Snippet: .. Real-time PCR was performed on a Roche LightCycler with a SYBR Green kit by using cDNA synthesized from total RNA isolated with TRIzol Reagent and further purified with the Qiagen RNeasy Mini kit. .. Serial dilutions of cDNA template (0.1, 0.01, 0.001, and 0.001) were amplified, and the cycle number vs. the log of the fluorescence measurement at the threshold was plotted to generate a standard curve for semiquantitation of mRNA expression.

Real-time Polymerase Chain Reaction:

Article Title: Assessing DNA methylation in the developing human intestinal epithelium: potential link to inflammatory bowel disease
Article Snippet: .. Complementary DNA corresponding to 5 ng RNA was used for real-time PCR using QuantiFast SYBR Green PCR Master Mix (Qiagen) on the 7500 Fast real-time PCR system, 7500 software v2.0.6 (Applied Biosciences by Thermo Fisher Scientific, Paisley, UK). .. Relative quantitation was calculated using the ΔΔC t method following normalization against GAPDH.

Article Title: Increased cholesterol 7?-hydroxylase expression and size of the bile acid pool in the lactating rat
Article Snippet: .. Real-time PCR was performed on a Roche LightCycler with a SYBR Green kit by using cDNA synthesized from total RNA isolated with TRIzol Reagent and further purified with the Qiagen RNeasy Mini kit. .. Serial dilutions of cDNA template (0.1, 0.01, 0.001, and 0.001) were amplified, and the cycle number vs. the log of the fluorescence measurement at the threshold was plotted to generate a standard curve for semiquantitation of mRNA expression.

Incubation:

Article Title: A ligand-directed divergent catalytic approach to establish structural and functional scaffold diversity
Article Snippet: .. After overnight incubation, cells were treated with 1.5 μM purmorphamine and the test compounds or DMSO for 48 h. Complementary DNA was prepared using FastLane Cell cDNA Kit (Qiagen) following the manufacturer's instructions. .. The expression of Hh target genes Ptch1 and Gli1 and the reference gene Gapdh gene was determined by means of quantitative PCR using the QuantiFast SYBR Green PCR Kit (Qiagen) and iQ 5 Real-Time PCR Detection System (Bio-rad) and the following oligonucleotides: Ptch1 : 5′-CTCTGGAGCAGATTTCCAAGG-3′ and 5′-TGCCGCAGTTCTTTTGAATG-3′, Gli1 : 5′-GGAAGTCCTATTCACGCCTTGA-3′ and 5′-CAACCTTCTTGCTCACACATGTAAG-3′, Gapdh : 5′-AGCCTCGTCCCG TAGACAAAAT-3′ and 5′-CCGTGAGTGGAGTCATACTGGA-3′ .

Polymerase Chain Reaction:

Article Title: Regulation of a Novel ?N-Catenin Splice Variant in Schizophrenic Smokers
Article Snippet: .. PCR amplification of cDNA was performed using 500 nM of primers and the Quantitect SYBR Green PCR Kit (Qiagen). .. PCR cycling conditions were 50°C for 2 min, 95°C for 15min, and 40 cycles of 94°C for 15 sec, 58°C for 30 sec, and 72°C for 30 sec.

Article Title: Assessing DNA methylation in the developing human intestinal epithelium: potential link to inflammatory bowel disease
Article Snippet: .. Complementary DNA corresponding to 5 ng RNA was used for real-time PCR using QuantiFast SYBR Green PCR Master Mix (Qiagen) on the 7500 Fast real-time PCR system, 7500 software v2.0.6 (Applied Biosciences by Thermo Fisher Scientific, Paisley, UK). .. Relative quantitation was calculated using the ΔΔC t method following normalization against GAPDH.

Plasmid Preparation:

Article Title: The Phosphorylation of Vinculin on Tyrosine Residues 100 and 1065, Mediated by Src Kinases, Affects Cell Spreading
Article Snippet: .. The complete cDNA encoding for chicken vinculin ( ) kindly provided by Susan W. Craig (John Hopkins University, Baltimore, MD) was subcloned in frame into the Eco RI site of a pQE-31 vector (QIAGEN), resulting in the addition of a 6-His tag to the carboxy-terminal end of vinculin. .. The gene was then subcloned into the Eco RI site of pcDNA3.1(+) (Invitrogen, San Diego, CA) for mammalian expression.

Software:

Article Title: Assessing DNA methylation in the developing human intestinal epithelium: potential link to inflammatory bowel disease
Article Snippet: .. Complementary DNA corresponding to 5 ng RNA was used for real-time PCR using QuantiFast SYBR Green PCR Master Mix (Qiagen) on the 7500 Fast real-time PCR system, 7500 software v2.0.6 (Applied Biosciences by Thermo Fisher Scientific, Paisley, UK). .. Relative quantitation was calculated using the ΔΔC t method following normalization against GAPDH.

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    Qiagen cdna synthesis total rna
    Example of reticulocyte <t>cDNA</t> quality control. Agarose gel electrophoresis of RT ‐ PCR products (image inverted, black/white) obtained using reticulocyte cDNA from patients ( HA patient, N62, and HS patient, C14) and controls (healthy individuals: C1 and C2) and primers encoding sequences of the following genes: CD 45 ( PTPRC gene; 217 bp)—leukocyte marker; β‐globin ( HBB gene; 397 bp)—erythroid marker; integrin‐β3 ( ITGB 3 gene; 198 bp)—platelet marker. As the “low abundance” gene transcript PPARA (peroxisome proliferator activated receptor alpha; 324 bp) was chosen and was taken from the end of the reticulocyte cDNA library ( NCBI , Library 11923). For the loading control β‐actin primers ( ACTB gene; 479 bp) were used. All primer sequences used in this experiment are included in Supporting Information Table S2 . As a standard “GeneRuler 100 bp DNA Ladder” (Thermo Fisher Scientific, Waltham, MA, USA) was used. Results show no leukocyte or thrombocyte <t>RNA</t> contamination for all analysed cases. The lowest band corresponds to primers
    Cdna Synthesis Total Rna, supplied by Qiagen, used in various techniques. Bioz Stars score: 92/100, based on 465 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen cdna construction
    RT–PCR amplification products from different tissues using the primer set CCV-mut-F/-R. Lane 1: Epstein Barr virus–transformed B-lymphocytes cell <t>cDNA</t> preparations showing the 218 bp transcripts. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and the acceptor splice site: TTT CCT CCA G^G CTG CAC CA-3′. Lane 2: A 500 bp genomic product. Lane 3: Universal Human reference <t>RNA</t> (mixed carcinoma) shows the 324 bp product. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and acceptor splice site: CTC TCC CCA G^C CAG CGC CCT-3′. Lane 4: cDNA from a human fetal eye from DNase-treated RNA. A 324 bp product (arrow) is seen. Lane 5: cDNA from human lens epithelial cells (HLEpiC cat. No. 6554 lot no. 2531, ScienCell) not DNase treated. A genomic DNA (500 bp) and a 324 bp PCR product (arrow) are seen from the spliced RP1–140A9.1 . Lane 6: 100 bp ladder.
    Cdna Construction, supplied by Qiagen, used in various techniques. Bioz Stars score: 90/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen cdna sequencing rna
    Flowchart illustrating the workflow of this study. This flowchart shows the general pipeline of this <t>RNA-seq</t> study, starting from sample preparation and RNA extraction to <t>cDNA</t> sequencing and data analyses. The biological interpretation of expression count was possible through the functional categories clustering of expressed genes.
    Cdna Sequencing Rna, supplied by Qiagen, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cdna sequencing rna/product/Qiagen
    Average 90 stars, based on 6 article reviews
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    cdna sequencing rna - by Bioz Stars, 2020-10
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    Image Search Results


    Example of reticulocyte cDNA quality control. Agarose gel electrophoresis of RT ‐ PCR products (image inverted, black/white) obtained using reticulocyte cDNA from patients ( HA patient, N62, and HS patient, C14) and controls (healthy individuals: C1 and C2) and primers encoding sequences of the following genes: CD 45 ( PTPRC gene; 217 bp)—leukocyte marker; β‐globin ( HBB gene; 397 bp)—erythroid marker; integrin‐β3 ( ITGB 3 gene; 198 bp)—platelet marker. As the “low abundance” gene transcript PPARA (peroxisome proliferator activated receptor alpha; 324 bp) was chosen and was taken from the end of the reticulocyte cDNA library ( NCBI , Library 11923). For the loading control β‐actin primers ( ACTB gene; 479 bp) were used. All primer sequences used in this experiment are included in Supporting Information Table S2 . As a standard “GeneRuler 100 bp DNA Ladder” (Thermo Fisher Scientific, Waltham, MA, USA) was used. Results show no leukocyte or thrombocyte RNA contamination for all analysed cases. The lowest band corresponds to primers

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Efficient method for isolation of reticulocyte RNA from healthy individuals and hemolytic anaemia patients, et al. Efficient method for isolation of reticulocyte RNA from healthy individuals and hemolytic anaemia patients

    doi: 10.1111/jcmm.13951

    Figure Lengend Snippet: Example of reticulocyte cDNA quality control. Agarose gel electrophoresis of RT ‐ PCR products (image inverted, black/white) obtained using reticulocyte cDNA from patients ( HA patient, N62, and HS patient, C14) and controls (healthy individuals: C1 and C2) and primers encoding sequences of the following genes: CD 45 ( PTPRC gene; 217 bp)—leukocyte marker; β‐globin ( HBB gene; 397 bp)—erythroid marker; integrin‐β3 ( ITGB 3 gene; 198 bp)—platelet marker. As the “low abundance” gene transcript PPARA (peroxisome proliferator activated receptor alpha; 324 bp) was chosen and was taken from the end of the reticulocyte cDNA library ( NCBI , Library 11923). For the loading control β‐actin primers ( ACTB gene; 479 bp) were used. All primer sequences used in this experiment are included in Supporting Information Table S2 . As a standard “GeneRuler 100 bp DNA Ladder” (Thermo Fisher Scientific, Waltham, MA, USA) was used. Results show no leukocyte or thrombocyte RNA contamination for all analysed cases. The lowest band corresponds to primers

    Article Snippet: 2.4 RNA isolation and cDNA synthesis Total RNA was extracted from the reticulocyte‐rich suspension and collected using the miRNeasy Mini Kit (Qiagen, Hilden, Germany), according to the manufacturer's recommendation.

    Techniques: Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Marker, cDNA Library Assay

    RT–PCR amplification products from different tissues using the primer set CCV-mut-F/-R. Lane 1: Epstein Barr virus–transformed B-lymphocytes cell cDNA preparations showing the 218 bp transcripts. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and the acceptor splice site: TTT CCT CCA G^G CTG CAC CA-3′. Lane 2: A 500 bp genomic product. Lane 3: Universal Human reference RNA (mixed carcinoma) shows the 324 bp product. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and acceptor splice site: CTC TCC CCA G^C CAG CGC CCT-3′. Lane 4: cDNA from a human fetal eye from DNase-treated RNA. A 324 bp product (arrow) is seen. Lane 5: cDNA from human lens epithelial cells (HLEpiC cat. No. 6554 lot no. 2531, ScienCell) not DNase treated. A genomic DNA (500 bp) and a 324 bp PCR product (arrow) are seen from the spliced RP1–140A9.1 . Lane 6: 100 bp ladder.

    Journal: Molecular Vision

    Article Title: A splice-site variant in the lncRNA gene RP1-140A9.1 cosegregates in the large Volkmann cataract family

    doi:

    Figure Lengend Snippet: RT–PCR amplification products from different tissues using the primer set CCV-mut-F/-R. Lane 1: Epstein Barr virus–transformed B-lymphocytes cell cDNA preparations showing the 218 bp transcripts. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and the acceptor splice site: TTT CCT CCA G^G CTG CAC CA-3′. Lane 2: A 500 bp genomic product. Lane 3: Universal Human reference RNA (mixed carcinoma) shows the 324 bp product. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and acceptor splice site: CTC TCC CCA G^C CAG CGC CCT-3′. Lane 4: cDNA from a human fetal eye from DNase-treated RNA. A 324 bp product (arrow) is seen. Lane 5: cDNA from human lens epithelial cells (HLEpiC cat. No. 6554 lot no. 2531, ScienCell) not DNase treated. A genomic DNA (500 bp) and a 324 bp PCR product (arrow) are seen from the spliced RP1–140A9.1 . Lane 6: 100 bp ladder.

    Article Snippet: RNA isolation, cDNA construction, and qPCR Total RNA was extracted from Epstein Barr transformed B-lymphocytes [ ], whole venous blood was taken from five healthy and affected individuals individuals (later frozen) and from human eye [ ] using an RNeasy mini kit from Qiagen (Cat No./ID: 74,104; Hilden, Germany).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Transformation Assay, CTG Assay, Polymerase Chain Reaction

    Overexpression of Tmie causes increased bundle expression of Tmc1-GFP and Tmc2b-GFP. (A) Semi-quantitative PCR of tmie cDNA using primers within exon 4 to detect transcripts from both transgene and the endogenous gene. We generated cDNA from RNA extracted from 5 dpf whole larvae. Larvae carrying Tg(myo6b : tmie-p2A-NLS(mCherry)) had 6.5-fold more tmie transcript than wild type non-transgenic siblings. Products are from 40 cycles. Less PCR reaction (2.5x less) was loaded for gapdh to avoid saturation. (B) Confocal images of the lateral cristae in 4 dpf tmie ru1000 larvae co-expressing two transgenes, tmc1-GFP (upper panel) and tmie-p2A-NLS(mCherry) (lower panel). The p2A linker is a self-cleaving peptide that results in equimolar translation of Tmie and nuclear mCherry. (C) Plot of the integrated density of Tmc1-GFP fluorescence/crista in the ROI, expressed in arbitrary units; significance determined by one-way ANOVA. (D-E) Same as B-C except using tmc2b-GFP instead of tmc1-GFP . All statistics are mean ± SD. Scale bars are 5μm.

    Journal: PLoS Genetics

    Article Title: Subunits of the mechano-electrical transduction channel, Tmc1/2b, require Tmie to localize in zebrafish sensory hair cells

    doi: 10.1371/journal.pgen.1007635

    Figure Lengend Snippet: Overexpression of Tmie causes increased bundle expression of Tmc1-GFP and Tmc2b-GFP. (A) Semi-quantitative PCR of tmie cDNA using primers within exon 4 to detect transcripts from both transgene and the endogenous gene. We generated cDNA from RNA extracted from 5 dpf whole larvae. Larvae carrying Tg(myo6b : tmie-p2A-NLS(mCherry)) had 6.5-fold more tmie transcript than wild type non-transgenic siblings. Products are from 40 cycles. Less PCR reaction (2.5x less) was loaded for gapdh to avoid saturation. (B) Confocal images of the lateral cristae in 4 dpf tmie ru1000 larvae co-expressing two transgenes, tmc1-GFP (upper panel) and tmie-p2A-NLS(mCherry) (lower panel). The p2A linker is a self-cleaving peptide that results in equimolar translation of Tmie and nuclear mCherry. (C) Plot of the integrated density of Tmc1-GFP fluorescence/crista in the ROI, expressed in arbitrary units; significance determined by one-way ANOVA. (D-E) Same as B-C except using tmc2b-GFP instead of tmc1-GFP . All statistics are mean ± SD. Scale bars are 5μm.

    Article Snippet: cDNA generation by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and semi-quantitative PCR For and , we extracted total RNA using the RNeasy mini kit (Qiagen).

    Techniques: Over Expression, Expressing, Real-time Polymerase Chain Reaction, Generated, Transgenic Assay, Polymerase Chain Reaction, Fluorescence

    Flowchart illustrating the workflow of this study. This flowchart shows the general pipeline of this RNA-seq study, starting from sample preparation and RNA extraction to cDNA sequencing and data analyses. The biological interpretation of expression count was possible through the functional categories clustering of expressed genes.

    Journal: Frontiers in Microbiology

    Article Title: Faustovirus E12 Transcriptome Analysis Reveals Complex Splicing in Capsid Gene

    doi: 10.3389/fmicb.2018.02534

    Figure Lengend Snippet: Flowchart illustrating the workflow of this study. This flowchart shows the general pipeline of this RNA-seq study, starting from sample preparation and RNA extraction to cDNA sequencing and data analyses. The biological interpretation of expression count was possible through the functional categories clustering of expressed genes.

    Article Snippet: RNA Extraction and cDNA Sequencing RNA was extracted using the RNeasy mini kit (Cat No: 74104, Qiagen, France) according to the manufacturer’s instructions.

    Techniques: RNA Sequencing Assay, Sample Prep, RNA Extraction, Sequencing, Expressing, Functional Assay