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  • 99
    Thermo Fisher iscript cdna synthesis kit
    Iscript Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 646 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/iscript cdna synthesis kit/product/Thermo Fisher
    Average 99 stars, based on 646 article reviews
    Price from $9.99 to $1999.99
    iscript cdna synthesis kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Bio-Rad iscript cdna synthesis kit
    RNA immunoprecipitation of (TMG)-capped primary miRNAs (Pri-miRNAs) in quiescent human foreskin fibroblasts (HFFs) RT-PCR data shows the RNA immunoprecipitation of Pri-miR-34a and Pri-miR-3188 in quiescent HFFs (as well as the positive control snRNA U7), but not Pri-miR-423 using an antibody against (TMG)-capped RNAs. Total RNA was extracted using TRIzol Reagent, and 10 μg of RNA was diluted in NET-2 buffer, precleared and incubated with Protein G Sepharose 4 Fast Flow beads loaded with 15 μl of control antibody (Normal Rabbit Serum, EMD-Millipore) or with antibody recognizing the (TMG)-cap (Anti-m3G-cap, rabbit polyclonal, Synaptic Systems). Beads were rinsed five times with NET-2 buffer and were resuspended in G-50 buffer. RNA was extracted from the beads by phenol-chloroform-isoamyl alcohol extraction and resuspended in 20 μl of nuclease-free water. Immunoprecipitated tri-methylated capped RNA was converted to <t>cDNA</t> using <t>iScript</t> cDNA synthesis kit (Bio-Rad), followed by RT-PCR, and visualized after gel electrophoresis.
    Iscript Cdna Synthesis Kit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 98645 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/iscript cdna synthesis kit/product/Bio-Rad
    Average 99 stars, based on 98645 article reviews
    Price from $9.99 to $1999.99
    iscript cdna synthesis kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    91
    Bio-Rad iscript gdna clear cdna synthesis kit
    RNA immunoprecipitation of (TMG)-capped primary miRNAs (Pri-miRNAs) in quiescent human foreskin fibroblasts (HFFs) RT-PCR data shows the RNA immunoprecipitation of Pri-miR-34a and Pri-miR-3188 in quiescent HFFs (as well as the positive control snRNA U7), but not Pri-miR-423 using an antibody against (TMG)-capped RNAs. Total RNA was extracted using TRIzol Reagent, and 10 μg of RNA was diluted in NET-2 buffer, precleared and incubated with Protein G Sepharose 4 Fast Flow beads loaded with 15 μl of control antibody (Normal Rabbit Serum, EMD-Millipore) or with antibody recognizing the (TMG)-cap (Anti-m3G-cap, rabbit polyclonal, Synaptic Systems). Beads were rinsed five times with NET-2 buffer and were resuspended in G-50 buffer. RNA was extracted from the beads by phenol-chloroform-isoamyl alcohol extraction and resuspended in 20 μl of nuclease-free water. Immunoprecipitated tri-methylated capped RNA was converted to <t>cDNA</t> using <t>iScript</t> cDNA synthesis kit (Bio-Rad), followed by RT-PCR, and visualized after gel electrophoresis.
    Iscript Gdna Clear Cdna Synthesis Kit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 406 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/iscript gdna clear cdna synthesis kit/product/Bio-Rad
    Average 91 stars, based on 406 article reviews
    Price from $9.99 to $1999.99
    iscript gdna clear cdna synthesis kit - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

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    RNA immunoprecipitation of (TMG)-capped primary miRNAs (Pri-miRNAs) in quiescent human foreskin fibroblasts (HFFs) RT-PCR data shows the RNA immunoprecipitation of Pri-miR-34a and Pri-miR-3188 in quiescent HFFs (as well as the positive control snRNA U7), but not Pri-miR-423 using an antibody against (TMG)-capped RNAs. Total RNA was extracted using TRIzol Reagent, and 10 μg of RNA was diluted in NET-2 buffer, precleared and incubated with Protein G Sepharose 4 Fast Flow beads loaded with 15 μl of control antibody (Normal Rabbit Serum, EMD-Millipore) or with antibody recognizing the (TMG)-cap (Anti-m3G-cap, rabbit polyclonal, Synaptic Systems). Beads were rinsed five times with NET-2 buffer and were resuspended in G-50 buffer. RNA was extracted from the beads by phenol-chloroform-isoamyl alcohol extraction and resuspended in 20 μl of nuclease-free water. Immunoprecipitated tri-methylated capped RNA was converted to cDNA using iScript cDNA synthesis kit (Bio-Rad), followed by RT-PCR, and visualized after gel electrophoresis.

    Journal: Bio-protocol

    Article Title: Immunoprecipitation of Tri-methylated Capped RNA

    doi: 10.21769/BioProtoc.2717

    Figure Lengend Snippet: RNA immunoprecipitation of (TMG)-capped primary miRNAs (Pri-miRNAs) in quiescent human foreskin fibroblasts (HFFs) RT-PCR data shows the RNA immunoprecipitation of Pri-miR-34a and Pri-miR-3188 in quiescent HFFs (as well as the positive control snRNA U7), but not Pri-miR-423 using an antibody against (TMG)-capped RNAs. Total RNA was extracted using TRIzol Reagent, and 10 μg of RNA was diluted in NET-2 buffer, precleared and incubated with Protein G Sepharose 4 Fast Flow beads loaded with 15 μl of control antibody (Normal Rabbit Serum, EMD-Millipore) or with antibody recognizing the (TMG)-cap (Anti-m3G-cap, rabbit polyclonal, Synaptic Systems). Beads were rinsed five times with NET-2 buffer and were resuspended in G-50 buffer. RNA was extracted from the beads by phenol-chloroform-isoamyl alcohol extraction and resuspended in 20 μl of nuclease-free water. Immunoprecipitated tri-methylated capped RNA was converted to cDNA using iScript cDNA synthesis kit (Bio-Rad), followed by RT-PCR, and visualized after gel electrophoresis.

    Article Snippet: Protein G Sepharose® 4 Fast Flow Beads (GE Healthcare, catalog number: 17061801) Normal rabbit serum (control, EMD Millipore, catalog number: NS01L-1ML) Sodium chloride (NaCl) (Fisher Scientific, catalog number: S671-3) NP-40 (Thermo Fisher Scientific, catalog number: 28324) Tris base (Fisher Scientific, catalog number: BP152-5) Hydrochloric acid (HCl) (VWR, catalog number: BDH7204-1) RNasin™ Plus RNase inhibitor (Promega, catalog number: N2611) NaOAc (AMRESCO, catalog number: 0602) Ethylenediaminetetraacetate acid (EDTA), pH 8 (Thermo Fisher Scientific, catalog number: AM9260G) Ethylenediaminetetraacetate acid (EDTA) (AMRESCO, catalog number: 0105) Sodium dodecyl sulfate (SDS) (Thermo Fisher Scientific, catalog number: AM9822) Phenol/Chloroform/Isoamyl Alcohol; 125:24:1 mixture, pH 4.5 (Thermo Fisher Scientific, catalog number: AM9720) Agarose LE (Denville Scientific, catalog number: CA3510-8) iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, catalog number: 1708891) Sso Advanced™ Universal SYBR® Green Supermix (Bio-Rad Laboratories, catalog number: 1725274) PARIS™ Kit (Thermo Fisher Scientific, catalog number: AM1921) Boric acid (Fisher Scientific, catalog number: A73-1) NET-2 Buffer (see Recipes) G-50 Buffer (see Recipes) 1× TBE (see Recipes)

    Techniques: Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction, Positive Control, Incubation, Flow Cytometry, Methylation, Nucleic Acid Electrophoresis

    BdGCN5 splice variants differ in their conserved domain composition and absolute transcript abundance. (A) Genomic representation of GCN5 alternative transcripts. Exons are depicted as black boxes, UTRs are depicted as white boxes. Forward (F) and reverse (R) primers used for qPCR are presented at their appropriate locations. Black arrows represent alternative splice sites. Scale bar = 1,000 bps. (B) Predicted protein product representation of BdGCN5 variants. Black boxes represent conserved protein domains. NLS, nuclear localization signal; HAT, histone acetyltransferase domain; and BROMO, Bromodomain. (C) Confirmation of specific detection of BdGCN5 splice variants through qPCR, using the primer pairs depicted in (A) , MW, molecular weight marker. (D) Absolute RNA quantification of each BdGCN5 variant is presented as an absolute value in mol/μl of input cDNA (the cDNA reaction contained 75 ng/μl of input RNA from aerial tissue extractions), as well as the percentage of the total BdGCN5 transcript pool in each tested accession.

    Journal: Frontiers in Plant Science

    Article Title: Diversification of the Histone Acetyltransferase GCN5 through Alternative Splicing in Brachypodium distachyon

    doi: 10.3389/fpls.2017.02176

    Figure Lengend Snippet: BdGCN5 splice variants differ in their conserved domain composition and absolute transcript abundance. (A) Genomic representation of GCN5 alternative transcripts. Exons are depicted as black boxes, UTRs are depicted as white boxes. Forward (F) and reverse (R) primers used for qPCR are presented at their appropriate locations. Black arrows represent alternative splice sites. Scale bar = 1,000 bps. (B) Predicted protein product representation of BdGCN5 variants. Black boxes represent conserved protein domains. NLS, nuclear localization signal; HAT, histone acetyltransferase domain; and BROMO, Bromodomain. (C) Confirmation of specific detection of BdGCN5 splice variants through qPCR, using the primer pairs depicted in (A) , MW, molecular weight marker. (D) Absolute RNA quantification of each BdGCN5 variant is presented as an absolute value in mol/μl of input cDNA (the cDNA reaction contained 75 ng/μl of input RNA from aerial tissue extractions), as well as the percentage of the total BdGCN5 transcript pool in each tested accession.

    Article Snippet: RNA extractions were performed using EZ-10 Spin Column Plant RNA Mini-Preps Kit (Bio Basic Inc.). cDNA synthesis was performed using iScript Advanced cDNA Synthesis Kit (BioRad Laboratories Inc.). qPCR was performed using Green-2-Go qPCR Mastermix (Bio Basic Inc.) and gene-specific primers in a CFX Connect™ (BioRad Laboratories Inc.).

    Techniques: Real-time Polymerase Chain Reaction, HAT Assay, Molecular Weight, Marker, Variant Assay

    Breast tumor kinase expression in breast cancer cell lines. (a) Total RNA was isolated from the breast cancer cell lines using the RNeasy kit (Qiagen), and the cDNA was generated using iScript cDNA synthesis (Bio-Rad). The cDNA was amplified using primers specifically for breast tumor kinase (Brk) or β-actin. The relative level of Brk mRNA was determined using densitometric analysis and was normalized to the amount of β-actin. The level of Brk mRNA in the MDA-MB-468 cell line was set to 1; numbers indicate the fold increase compared with the 468 cells. (b) Whole-cell lysates were prepared from the breast cancer cell lines. Lysates were immunoblotted with anti-Brk antibody (top) and with anti-Ran antibody (bottom). The relative expression of Brk protein was determined using densitometric analysis and was normalized to the amount of Ran. The expression of Brk protein in the MDA-MB-468 cell line was set to 1; numbers indicate the fold increase compared with the 468 cells.

    Journal: Breast Cancer Research : BCR

    Article Title: Signal transducer and activator of transcription 5b: a new target of breast tumor kinase/protein tyrosine kinase 6

    doi: 10.1186/bcr1794

    Figure Lengend Snippet: Breast tumor kinase expression in breast cancer cell lines. (a) Total RNA was isolated from the breast cancer cell lines using the RNeasy kit (Qiagen), and the cDNA was generated using iScript cDNA synthesis (Bio-Rad). The cDNA was amplified using primers specifically for breast tumor kinase (Brk) or β-actin. The relative level of Brk mRNA was determined using densitometric analysis and was normalized to the amount of β-actin. The level of Brk mRNA in the MDA-MB-468 cell line was set to 1; numbers indicate the fold increase compared with the 468 cells. (b) Whole-cell lysates were prepared from the breast cancer cell lines. Lysates were immunoblotted with anti-Brk antibody (top) and with anti-Ran antibody (bottom). The relative expression of Brk protein was determined using densitometric analysis and was normalized to the amount of Ran. The expression of Brk protein in the MDA-MB-468 cell line was set to 1; numbers indicate the fold increase compared with the 468 cells.

    Article Snippet: The cDNA was generated using iScript cDNA synthesis (Bio-Rad).

    Techniques: Expressing, Isolation, Generated, Amplification, Multiple Displacement Amplification

    Induction of autophagic genes expression following 4 weeks of nitrite therapy in CHF mice. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of autophagic markers beclin‐1 (A), ATG ‐5 (B), and ATG ‐7 (C). The number in the circle inside the bar denotes the number of animals used. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). ATG indicates autophagy related gene; CHF, chronic heart failure.

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

    Article Title: Nitrite Therapy Ameliorates Myocardial Dysfunction via H2S and Nuclear Factor‐Erythroid 2‐Related Factor 2 (Nrf2)‐Dependent Signaling in Chronic Heart Failure

    doi: 10.1161/JAHA.116.003551

    Figure Lengend Snippet: Induction of autophagic genes expression following 4 weeks of nitrite therapy in CHF mice. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of autophagic markers beclin‐1 (A), ATG ‐5 (B), and ATG ‐7 (C). The number in the circle inside the bar denotes the number of animals used. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). ATG indicates autophagy related gene; CHF, chronic heart failure.

    Article Snippet: One microgram of RNA was transcribed using an I‐script cDNA synthesis kit from Bio‐Rad.

    Techniques: Expressing, Mouse Assay, Software

    Effects of nitrite therapy on hypertrophic gene expression in ischemia‐induced CHF mice. Nitrite (100 mg/L) was given in the drinking water during reperfusion. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of ANP (A), BNP (B), and Myh7 (C) using TaqMan PCR assay system. The number inside the bar denotes the number of animals used per experiment. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). ANP indicates atrial natriuretic peptide; BNP, brain natriuretic peptide; Myh7, myosin heavy chain beta; PCR, polymerase chain reaction.

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

    Article Title: Nitrite Therapy Ameliorates Myocardial Dysfunction via H2S and Nuclear Factor‐Erythroid 2‐Related Factor 2 (Nrf2)‐Dependent Signaling in Chronic Heart Failure

    doi: 10.1161/JAHA.116.003551

    Figure Lengend Snippet: Effects of nitrite therapy on hypertrophic gene expression in ischemia‐induced CHF mice. Nitrite (100 mg/L) was given in the drinking water during reperfusion. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of ANP (A), BNP (B), and Myh7 (C) using TaqMan PCR assay system. The number inside the bar denotes the number of animals used per experiment. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). ANP indicates atrial natriuretic peptide; BNP, brain natriuretic peptide; Myh7, myosin heavy chain beta; PCR, polymerase chain reaction.

    Article Snippet: One microgram of RNA was transcribed using an I‐script cDNA synthesis kit from Bio‐Rad.

    Techniques: Expressing, Mouse Assay, Aqueous Normal-phase Chromatography, Polymerase Chain Reaction, Software

    Effects of nitrite on cardiac antioxidant gene and protein levels in CHF mice. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of SOD 1 (A), catalase (B), and GPX (C) using TaqMan PCR assay system. (D) represents proteins levels of SOD 1, catalase, and GPX , and (E), (F), and (G) represent the quantitation of the blots in (D). The number in the circle inside the bar denotes the number of animals used. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). CHF indicates chronic heart failure; GPX, glutathione peroxidase; PCR, polymerase chain reaction; SOD1, superoxide dismutase 1.

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

    Article Title: Nitrite Therapy Ameliorates Myocardial Dysfunction via H2S and Nuclear Factor‐Erythroid 2‐Related Factor 2 (Nrf2)‐Dependent Signaling in Chronic Heart Failure

    doi: 10.1161/JAHA.116.003551

    Figure Lengend Snippet: Effects of nitrite on cardiac antioxidant gene and protein levels in CHF mice. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of SOD 1 (A), catalase (B), and GPX (C) using TaqMan PCR assay system. (D) represents proteins levels of SOD 1, catalase, and GPX , and (E), (F), and (G) represent the quantitation of the blots in (D). The number in the circle inside the bar denotes the number of animals used. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). CHF indicates chronic heart failure; GPX, glutathione peroxidase; PCR, polymerase chain reaction; SOD1, superoxide dismutase 1.

    Article Snippet: One microgram of RNA was transcribed using an I‐script cDNA synthesis kit from Bio‐Rad.

    Techniques: Mouse Assay, Polymerase Chain Reaction, Quantitation Assay, Software

    TSC-22 induces p53 expression. ( A ) TSC-22 and p53 cDNA constructs were cotransformed into EGY48 yeast cells to test for protein–protein interaction within the yeast two-hybrid system. Transformants were assayed for their ability to grow on medium lacking leucine (left) and for β-galactoside expression (right). ( B ) HeLa cells and Caski cells were infected with Ad- TSC-22 for the indicated times. Protein levels were analyzed by Western blot with the DO-1 antibody against p53, the anti-TSC-22 antibody, and the anti-β-actin antibody as a loading control. ( C ) HeLa cells were transfected with 1 µg of Flag-TSC-22 expression vector. 24 h post-transfection, p53, Puma, p21, and TSC22 expression were analyzed by Western blotting and semi-quantitative RT-PCR with protein and total RNA obtained from each cell line. ( D ) HeLa cells stably expressing shRNA specific for TSC-22 and non-targeting control shRNA were analyzed to determine the protein and mRNA expression levels of p53 and Puma. ( E ) Luciferase activity of the p53RE (responsible element)-driven promoter were assessed with transfection of the indicated amount of Flag-TSC-22 plasmid in HeLa cells (upper panel). Activity of the p53RE-promoter was assessed in sh-con and sh-TSC-22 expressing HeLa cells (lower panel) by luciferase assays. ( F ) One µg of Flag-TSC-22 or Flag-mock vector was transfected into p53 +/+ or p53 −/− HCT116 cells. At 48 h post-transfection, cell lysates were analyzed by Western blotting with the indicated antibodies. ( G ) Stability of p53 protein was assessed in HeLa cells infected with Ad- TSC-22 or Ad- LacZ . 24 h after infection, cells were treated with cycloheximide (50 µg/mL) for the indicated periods of time. Cell lysates were analyzed by Western blotting with anti-p53 antibody (DO-1) with β-actin as a loading control.

    Journal: PLoS ONE

    Article Title: Crucial Role of TSC-22 in Preventing the Proteasomal Degradation of p53 in Cervical Cancer

    doi: 10.1371/journal.pone.0042006

    Figure Lengend Snippet: TSC-22 induces p53 expression. ( A ) TSC-22 and p53 cDNA constructs were cotransformed into EGY48 yeast cells to test for protein–protein interaction within the yeast two-hybrid system. Transformants were assayed for their ability to grow on medium lacking leucine (left) and for β-galactoside expression (right). ( B ) HeLa cells and Caski cells were infected with Ad- TSC-22 for the indicated times. Protein levels were analyzed by Western blot with the DO-1 antibody against p53, the anti-TSC-22 antibody, and the anti-β-actin antibody as a loading control. ( C ) HeLa cells were transfected with 1 µg of Flag-TSC-22 expression vector. 24 h post-transfection, p53, Puma, p21, and TSC22 expression were analyzed by Western blotting and semi-quantitative RT-PCR with protein and total RNA obtained from each cell line. ( D ) HeLa cells stably expressing shRNA specific for TSC-22 and non-targeting control shRNA were analyzed to determine the protein and mRNA expression levels of p53 and Puma. ( E ) Luciferase activity of the p53RE (responsible element)-driven promoter were assessed with transfection of the indicated amount of Flag-TSC-22 plasmid in HeLa cells (upper panel). Activity of the p53RE-promoter was assessed in sh-con and sh-TSC-22 expressing HeLa cells (lower panel) by luciferase assays. ( F ) One µg of Flag-TSC-22 or Flag-mock vector was transfected into p53 +/+ or p53 −/− HCT116 cells. At 48 h post-transfection, cell lysates were analyzed by Western blotting with the indicated antibodies. ( G ) Stability of p53 protein was assessed in HeLa cells infected with Ad- TSC-22 or Ad- LacZ . 24 h after infection, cells were treated with cycloheximide (50 µg/mL) for the indicated periods of time. Cell lysates were analyzed by Western blotting with anti-p53 antibody (DO-1) with β-actin as a loading control.

    Article Snippet: First strand cDNA was prepared from total RNA and oligo dT using I Script cDNA synthesis kit (Bio-Rad, USA).

    Techniques: Expressing, Construct, Infection, Western Blot, Transfection, Plasmid Preparation, Quantitative RT-PCR, Stable Transfection, shRNA, Luciferase, Activity Assay