rt pcr  (Thermo Fisher)

 
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    Name:
    SuperScript First Strand Synthesis System for RT PCR
    Description:
    The SuperScript First Strand Synthesis System for RT PCR is optimized to synthesize first strand cDNA from purified poly A or total RNA The system can be used with as little as 1 ng or as much as 5 µg of total RNA After synthesis the cDNA can be amplified with specific primers by PCR without intermediate organic extractions or ethanol precipitations In conjunction with PCR the system can be used to detect the presence of rare messages to quantitate the amount of specific mRNA from small numbers of cells or to clone specific cDNAs without constructing an entire cDNA library The system is flexible allowing the use of any PCR enzyme Combine with AccuPrime Taq DNA Polymerase or Platinum Taq DNA Polymerase for higher specificity PCR or with AccuPrime Pfx DNA Polymerase for high fidelity cloning applications SuperScript II RTThe first strand cDNA synthesis reaction is catalyzed by SuperScript II Reverse Transcriptase RT This enzyme has been engineered to reduce the RNase H activity that degrades mRNA during the first strand reaction resulting in greater full length cDNA synthesis and higher yields of first strand cDNA than obtained with RNase H RTs Because SuperScript II RT is not inhibited significantly by ribosomal and transfer RNA it may be used effectively to synthesize first strand cDNA from a total RNA preparation The enzyme exhibits increased thermal stability and may be used at temperatures up to 50°C Using the SuperScript First Strand Synthesis System for RT PCRThis system has been optimized to synthesize first strand cDNA from varying amounts of starting material The SuperScript II RT concentration has been lowered and RNaseOUT Recombinant RNase Inhibitor has been added to the system as part of this optimization process Additionally reaction conditions have been modified to further increase the sensitivity of the system Using the kit you synthesize first strand cDNA using either total RNA or poly A selected RNA primed with oligo dT random primers or a gene specific primer Then you perform PCR in a separate tube using primers specific for the gene of interest
    Catalog Number:
    11904018
    Price:
    None
    Applications:
    PCR & Real-Time PCR|Reverse Transcription
    Category:
    Kits and Assays
    Buy from Supplier


    Structured Review

    Thermo Fisher rt pcr
    The SuperScript First Strand Synthesis System for RT PCR is optimized to synthesize first strand cDNA from purified poly A or total RNA The system can be used with as little as 1 ng or as much as 5 µg of total RNA After synthesis the cDNA can be amplified with specific primers by PCR without intermediate organic extractions or ethanol precipitations In conjunction with PCR the system can be used to detect the presence of rare messages to quantitate the amount of specific mRNA from small numbers of cells or to clone specific cDNAs without constructing an entire cDNA library The system is flexible allowing the use of any PCR enzyme Combine with AccuPrime Taq DNA Polymerase or Platinum Taq DNA Polymerase for higher specificity PCR or with AccuPrime Pfx DNA Polymerase for high fidelity cloning applications SuperScript II RTThe first strand cDNA synthesis reaction is catalyzed by SuperScript II Reverse Transcriptase RT This enzyme has been engineered to reduce the RNase H activity that degrades mRNA during the first strand reaction resulting in greater full length cDNA synthesis and higher yields of first strand cDNA than obtained with RNase H RTs Because SuperScript II RT is not inhibited significantly by ribosomal and transfer RNA it may be used effectively to synthesize first strand cDNA from a total RNA preparation The enzyme exhibits increased thermal stability and may be used at temperatures up to 50°C Using the SuperScript First Strand Synthesis System for RT PCRThis system has been optimized to synthesize first strand cDNA from varying amounts of starting material The SuperScript II RT concentration has been lowered and RNaseOUT Recombinant RNase Inhibitor has been added to the system as part of this optimization process Additionally reaction conditions have been modified to further increase the sensitivity of the system Using the kit you synthesize first strand cDNA using either total RNA or poly A selected RNA primed with oligo dT random primers or a gene specific primer Then you perform PCR in a separate tube using primers specific for the gene of interest
    https://www.bioz.com/result/rt pcr/product/Thermo Fisher
    Average 97 stars, based on 17152 article reviews
    Price from $9.99 to $1999.99
    rt pcr - by Bioz Stars, 2021-02
    97/100 stars

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    Related Articles

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Upregulation of SQSTM1/p62 contributes to nickel-induced malignant transformation of human bronchial epithelial cells
    Article Snippet: .. RT-PCR Cells were exposed to nickel for the indicated time points, and then 5.0 μg total RNA was used for first-strand cDNA synthesis with oligodT (20) primer by SuperScriptTM First-Strand Synthesis system (Invitrogen, 11904018). .. The SQSTM1 and TNF were monitored by PCR.

    Article Title: M-CSF Induces Vascular Endothelial Growth Factor Production and Angiogenic Activity From Human Monocytes
    Article Snippet: .. RNA integrity was verified on a 1.5% agarose gel to detect 18S and 28S ribosomal RNA bands at 0.7 and 1.5 kDa, respectively. cDNA was synthesized from monocyte total cellular RNA using the SuperScript First-Strand Synthesis System for RT-PCR Kit (Life Technologies). .. The VEGF forward primer, VEGF reverse primer, and the VEGF probe with TAMRA quencher (PE Applied Biosystems) for VEGF mRNA were designed using Primer Express version 1.0 software (ABI PRISM, PerkinElmer, Branchburg, NJ) based on the human VEGF sequence obtained from PubMed (accession no. NM 003376).

    Article Title: Tropomyosin 3.1 Association With Actin Stress Fibers is Required for Lens Epithelial to Mesenchymal Transition
    Article Snippet: .. Reverse transcription was performed on equal amounts of total RNA for each sample using Superscript III First-Strand Synthesis System for RT-PCR Kit (Thermo Fisher Scientific). .. Semi-quantitative RT-PCR was performed using Hot-Start Taq Blue Mastermix (Thomas Scientific, Swedesboro, NJ, USA) on equivalent amounts of cDNA in a 20 µL reaction and using previously validated mouse Tpm primers.

    Article Title: Effects of the Sri Lankan medicinal plant, Salacia reticulata, in rheumatoid arthritis
    Article Snippet: .. The Single-strand cDNA was prepared from 1 μg of total RNA using the SuperScript First-Strand Synthesis System for RT-PCR (Invitrogen, Tokyo, Japan). .. Amplification was performed in 10 μl of reaction mixture using EX taq (TAKARA BIO INC, Shiga, Japan).

    Article Title: Tropomyosin 3.5 protects the F-actin networks required for tissue biomechanical properties
    Article Snippet: .. Reverse transcription was performed using the SuperscriptTM III First-Strand Synthesis System for RT-PCR kit (Thermo Fisher Scientific) from an equal amount of total RNA for each sample. .. PCR was performed with the same amount of cDNA from each sample in a 20 µl volume reaction.

    Real-time Polymerase Chain Reaction:

    Article Title: Bisphosphonates and statins inhibit expression and secretion of MIP-1α via suppression of Ras/MEK/ERK/AML-1A and Ras/PI3K/Akt/AML-1A pathways
    Article Snippet: .. One microgram of purified total RNA was used for the real-time PCR analysis with the SuperScript First-Strand Synthesis System (Invitrogen). cDNA was subjected to quantitative real-time PCR by using SYBR Premix Ex Taq (Takara Biomedical; Siga, Japan) and the Thermal Cycler Dice Real Time system (Takara Biomedical) in a 96-well plate according to the manufacturer’s instructions. .. The PCR conditions for glyceraldehyde-3-phosphate dehydrogenase (GAPDH), MIP-1α, AML-1A, CREB, NF-AT, and C/ERBβ were 94°C for 2 min; followed by 40 cycles of 94°C for 0.5 min, 50°C for 0.5 min, and 72°C for 0.5 min.

    Synthesized:

    Article Title: M-CSF Induces Vascular Endothelial Growth Factor Production and Angiogenic Activity From Human Monocytes
    Article Snippet: .. RNA integrity was verified on a 1.5% agarose gel to detect 18S and 28S ribosomal RNA bands at 0.7 and 1.5 kDa, respectively. cDNA was synthesized from monocyte total cellular RNA using the SuperScript First-Strand Synthesis System for RT-PCR Kit (Life Technologies). .. The VEGF forward primer, VEGF reverse primer, and the VEGF probe with TAMRA quencher (PE Applied Biosystems) for VEGF mRNA were designed using Primer Express version 1.0 software (ABI PRISM, PerkinElmer, Branchburg, NJ) based on the human VEGF sequence obtained from PubMed (accession no. NM 003376).

    Agarose Gel Electrophoresis:

    Article Title: M-CSF Induces Vascular Endothelial Growth Factor Production and Angiogenic Activity From Human Monocytes
    Article Snippet: .. RNA integrity was verified on a 1.5% agarose gel to detect 18S and 28S ribosomal RNA bands at 0.7 and 1.5 kDa, respectively. cDNA was synthesized from monocyte total cellular RNA using the SuperScript First-Strand Synthesis System for RT-PCR Kit (Life Technologies). .. The VEGF forward primer, VEGF reverse primer, and the VEGF probe with TAMRA quencher (PE Applied Biosystems) for VEGF mRNA were designed using Primer Express version 1.0 software (ABI PRISM, PerkinElmer, Branchburg, NJ) based on the human VEGF sequence obtained from PubMed (accession no. NM 003376).

    Purification:

    Article Title: Bisphosphonates and statins inhibit expression and secretion of MIP-1α via suppression of Ras/MEK/ERK/AML-1A and Ras/PI3K/Akt/AML-1A pathways
    Article Snippet: .. One microgram of purified total RNA was used for the real-time PCR analysis with the SuperScript First-Strand Synthesis System (Invitrogen). cDNA was subjected to quantitative real-time PCR by using SYBR Premix Ex Taq (Takara Biomedical; Siga, Japan) and the Thermal Cycler Dice Real Time system (Takara Biomedical) in a 96-well plate according to the manufacturer’s instructions. .. The PCR conditions for glyceraldehyde-3-phosphate dehydrogenase (GAPDH), MIP-1α, AML-1A, CREB, NF-AT, and C/ERBβ were 94°C for 2 min; followed by 40 cycles of 94°C for 0.5 min, 50°C for 0.5 min, and 72°C for 0.5 min.

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  • 90
    Thermo Fisher quantitative real time reverse transcription polymerase chain reaction qrt pcr
    Knockdown or suppression of p65 reduces GLS1 expression in HCC cells. ( A ) Western blot shows the knockdown of c-Myc without interfering with GLS1 expression in HepG2 and Hep3B cells. ( B ) Western blot shows that p65 knockdown reduces GLS1 protein levels in both HepG2 and Hep3B cells. ( C ) <t>qRT-PCR</t> indicates that p65 knockdown reduces GLS1 mRNA levels in both HepG2 and Hep3B cells. ** p
    Quantitative Real Time Reverse Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time reverse transcription polymerase chain reaction qrt pcr/product/Thermo Fisher
    Average 90 stars, based on 53 article reviews
    Price from $9.99 to $1999.99
    quantitative real time reverse transcription polymerase chain reaction qrt pcr - by Bioz Stars, 2021-02
    90/100 stars
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    89
    Thermo Fisher real time pcr rt pcr analysis
    Expression of HCO − 3 transporters in AE3-null hearts . <t>RT-PCR</t> analysis of <t>cDNA</t> generated from total RNA of WT and AE3-null (KO) hearts was carried out to determine mRNA levels the Slc4a4 and Slc4a7 Na + /HCO − 3 cotransporters and Slc26a6 , encoding the anion exchanger PAT1. When normalized to Gapdh levels, expression of Slc4a4 was downregulated in AE3-null hearts (A) , while Slc4a7 (B) and Sl26a6 (C) levels showed no change. Total cardiac homogenates of WT and KO hearts were subjected to immunoblot and densitometric analyses as described in Figure 3 ; results show that NBCe1 protein expression, when normalized to levels of sarcomeric actin (s.actin) was reduced in AE3-null hearts (D,E) . Values shown are mean ± S.E. n = at least 10 mice of each genotype for RT-PCR analysis and at least 7 mice of each genotype for immunoblot analysis. * p
    Real Time Pcr Rt Pcr Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 175 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time pcr rt pcr analysis/product/Thermo Fisher
    Average 89 stars, based on 175 article reviews
    Price from $9.99 to $1999.99
    real time pcr rt pcr analysis - by Bioz Stars, 2021-02
    89/100 stars
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    88
    Thermo Fisher semi quantitative reverse transcriptase polymerase chain reaction rt pcr total rna
    FHC is required for caffeine modulation of H460 proliferation. (A) Real-time <t>PCR</t> analysis of FHC mRNA amounts was performed on total <t>RNA</t> from H460 siFHC and H460 siRNA cells treated with the indicated doses of caffeine. Results are representative of two different experiments (* p
    Semi Quantitative Reverse Transcriptase Polymerase Chain Reaction Rt Pcr Total Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/semi quantitative reverse transcriptase polymerase chain reaction rt pcr total rna/product/Thermo Fisher
    Average 88 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    semi quantitative reverse transcriptase polymerase chain reaction rt pcr total rna - by Bioz Stars, 2021-02
    88/100 stars
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    Image Search Results


    Knockdown or suppression of p65 reduces GLS1 expression in HCC cells. ( A ) Western blot shows the knockdown of c-Myc without interfering with GLS1 expression in HepG2 and Hep3B cells. ( B ) Western blot shows that p65 knockdown reduces GLS1 protein levels in both HepG2 and Hep3B cells. ( C ) qRT-PCR indicates that p65 knockdown reduces GLS1 mRNA levels in both HepG2 and Hep3B cells. ** p

    Journal: OncoTargets and therapy

    Article Title: Nuclear factor-κB p65 regulates glutaminase 1 expression in human hepatocellular carcinoma

    doi: 10.2147/OTT.S167408

    Figure Lengend Snippet: Knockdown or suppression of p65 reduces GLS1 expression in HCC cells. ( A ) Western blot shows the knockdown of c-Myc without interfering with GLS1 expression in HepG2 and Hep3B cells. ( B ) Western blot shows that p65 knockdown reduces GLS1 protein levels in both HepG2 and Hep3B cells. ( C ) qRT-PCR indicates that p65 knockdown reduces GLS1 mRNA levels in both HepG2 and Hep3B cells. ** p

    Article Snippet: Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) Upon relative treatment, cells were lysed, and total RNA was isolated using TRIzol (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR

    Expression of HCO − 3 transporters in AE3-null hearts . RT-PCR analysis of cDNA generated from total RNA of WT and AE3-null (KO) hearts was carried out to determine mRNA levels the Slc4a4 and Slc4a7 Na + /HCO − 3 cotransporters and Slc26a6 , encoding the anion exchanger PAT1. When normalized to Gapdh levels, expression of Slc4a4 was downregulated in AE3-null hearts (A) , while Slc4a7 (B) and Sl26a6 (C) levels showed no change. Total cardiac homogenates of WT and KO hearts were subjected to immunoblot and densitometric analyses as described in Figure 3 ; results show that NBCe1 protein expression, when normalized to levels of sarcomeric actin (s.actin) was reduced in AE3-null hearts (D,E) . Values shown are mean ± S.E. n = at least 10 mice of each genotype for RT-PCR analysis and at least 7 mice of each genotype for immunoblot analysis. * p

    Journal: Frontiers in Physiology

    Article Title: Loss of the AE3 Cl−/HCO−3 exchanger in mice affects rate-dependent inotropy and stress-related AKT signaling in heart

    doi: 10.3389/fphys.2013.00399

    Figure Lengend Snippet: Expression of HCO − 3 transporters in AE3-null hearts . RT-PCR analysis of cDNA generated from total RNA of WT and AE3-null (KO) hearts was carried out to determine mRNA levels the Slc4a4 and Slc4a7 Na + /HCO − 3 cotransporters and Slc26a6 , encoding the anion exchanger PAT1. When normalized to Gapdh levels, expression of Slc4a4 was downregulated in AE3-null hearts (A) , while Slc4a7 (B) and Sl26a6 (C) levels showed no change. Total cardiac homogenates of WT and KO hearts were subjected to immunoblot and densitometric analyses as described in Figure 3 ; results show that NBCe1 protein expression, when normalized to levels of sarcomeric actin (s.actin) was reduced in AE3-null hearts (D,E) . Values shown are mean ± S.E. n = at least 10 mice of each genotype for RT-PCR analysis and at least 7 mice of each genotype for immunoblot analysis. * p

    Article Snippet: Real-time PCR (RT-PCR) analysis of cDNA was carried out using the Absolute Blue qPCR SYBR mix (Thermo Scientific; AB4219/B), the Opticon2 DNA Engine (MJ Research Inc.), and QuantiTect primer assays (Qiagen) targeted against mRNA for the following genes: Slc4a4 encoding NBCe1 (Primer assay: QT01053675), Slc4a7 encoding NBCn1 (Primer assay: QT01037036), and Slc26a6 encoding PAT1 (Primer assay: QT01661828).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Generated, Mouse Assay

    Validation of up-regulation of the carrier gene VMP1 by IL-4 in CLL. Two of the CLL patients, CLL06 and CLL14, were assayed by a semiquantitative RT-PCR assay. GAPDH was used as reference. The size of the molecular weight markers (M) is indicated to the left. The size of the expected PCR fragments were 606 bp for VMP1 and 464 bp for GAPDH.

    Journal: PLoS ONE

    Article Title: IL-4 Up-Regulates MiR-21 and the MiRNAs Hosted in the CLCN5 Gene in Chronic Lymphocytic Leukemia

    doi: 10.1371/journal.pone.0124936

    Figure Lengend Snippet: Validation of up-regulation of the carrier gene VMP1 by IL-4 in CLL. Two of the CLL patients, CLL06 and CLL14, were assayed by a semiquantitative RT-PCR assay. GAPDH was used as reference. The size of the molecular weight markers (M) is indicated to the left. The size of the expected PCR fragments were 606 bp for VMP1 and 464 bp for GAPDH.

    Article Snippet: Semiquantitative RT-PCR The levels of expression of VMP1 were measured using the GeneAmp RNA PCR Core Kit in a GeneAmp PCR System 9700 (both from Thermo Fisher Scientific), following the manufacturer’s instructions.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Molecular Weight, Polymerase Chain Reaction

    FHC is required for caffeine modulation of H460 proliferation. (A) Real-time PCR analysis of FHC mRNA amounts was performed on total RNA from H460 siFHC and H460 siRNA cells treated with the indicated doses of caffeine. Results are representative of two different experiments (* p

    Journal: PLoS ONE

    Article Title: Caffeine Positively Modulates Ferritin Heavy Chain Expression in H460 Cells: Effects on Cell Proliferation

    doi: 10.1371/journal.pone.0163078

    Figure Lengend Snippet: FHC is required for caffeine modulation of H460 proliferation. (A) Real-time PCR analysis of FHC mRNA amounts was performed on total RNA from H460 siFHC and H460 siRNA cells treated with the indicated doses of caffeine. Results are representative of two different experiments (* p

    Article Snippet: RNA extraction and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) Total RNA was extracted from H460 and SKOV3 cells with the TRizol RNA isolation system (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

    Techniques: Real-time Polymerase Chain Reaction

    FHC is required for caffeine modulation of SKOV3 cells proliferation. (A) Real-time PCR analysis of FHC mRNA amounts was performed on total RNA from SKOV3 pcDNA and from SKOV3 pcFHC cells. Results are representative of two different experiments (* p

    Journal: PLoS ONE

    Article Title: Caffeine Positively Modulates Ferritin Heavy Chain Expression in H460 Cells: Effects on Cell Proliferation

    doi: 10.1371/journal.pone.0163078

    Figure Lengend Snippet: FHC is required for caffeine modulation of SKOV3 cells proliferation. (A) Real-time PCR analysis of FHC mRNA amounts was performed on total RNA from SKOV3 pcDNA and from SKOV3 pcFHC cells. Results are representative of two different experiments (* p

    Article Snippet: RNA extraction and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) Total RNA was extracted from H460 and SKOV3 cells with the TRizol RNA isolation system (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

    Techniques: Real-time Polymerase Chain Reaction

    RNA extraction and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR)

    Journal: PLoS ONE

    Article Title: Caffeine Positively Modulates Ferritin Heavy Chain Expression in H460 Cells: Effects on Cell Proliferation

    doi: 10.1371/journal.pone.0163078

    Figure Lengend Snippet: RNA extraction and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR)

    Article Snippet: RNA extraction and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) Total RNA was extracted from H460 and SKOV3 cells with the TRizol RNA isolation system (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

    Techniques: RNA Extraction, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction