RT-PCR for gene expression analysis: PrimeScript High Fidelity RT-PCR Kit
PrimeScript Reverse Transcriptase (PrimeScript RT), included in the PrimeScript High Fidelity RT-PCR Kit, efficiently synthesizes cDNA thanks to its exceptionally strong strand-displacement activity. PrimeScript RT is a modified, recombinant RNase H Minus MMLV (Moloney Murine Leukemia Virus) reverse transcriptase that is robust, versatile, and well-suited for applications requiring full-length cDNA, such as RT-PCR gene expression studies. PrimeScript RT can be used for reverse transcription of virtually any RNA template, including templates that are GC-rich and/or have high levels of secondary structure. Because of the excellent extension capability of this enzyme, cDNA synthesis can be performed at 42°C, decreasing the risk of RNA degradation that often occurs during conventional reactions performed at higher temperatures.
PrimeScript Reverse Transcriptase (PrimeScript RT), included in the PrimeScript High Fidelity RT-PCR Kit, efficiently synthesizes cDNA thanks to its exceptionally strong strand-displacement activity. PrimeScript RT is a modified, recombinant RNase H Minus MMLV (Moloney Murine Leukemia Virus) reverse transcriptase that is robust, versatile, and well-suited for applications requiring full-length cDNA, such as RT-PCR gene expression studies. PrimeScript RT can be used for reverse transcription of virtually any RNA template, including templates that are GC-rich and/or have high levels of secondary structure. Because of the excellent extension capability of this enzyme, cDNA synthesis can be performed at 42°C, decreasing the risk of RNA degradation that often occurs during conventional reactions performed at higher temperatures.
Fast, accurate quantification
During RT-PCR gene expression studies, accurate reverse transcription and PCR amplification are essential. The PrimeScript High Fidelity RT-PCR Kit includes all of the reagents needed for high-fidelity amplification of cDNA from total RNA or mRNA, and is designed for two-step endpoint RT-PCR. This kit includes PrimeScript High Fidelity Reverse Transcriptase, a highly accurate MMLV reverse transcriptase, and PrimeSTAR Max DNA Polymerase, which has the highest fidelity of any commercially available PCR enzyme. PrimeSTAR Max DNA Polymerase is formulated as a premix and offers high amplification efficiency as well as compatibility with a wide range of template concentrations. As a result, this kit is well-suited for any cDNA studies requiring high fidelity, such as RT-PCR gene expression studies (including gene-splicing studies where alternative splicing products must be distinguished, or when discerning between closely related gene family members), cDNA library construction, and transcriptome analysis.
Proven performance
First introduced in Japan, PrimeScript RT is now available to scientists worldwide. Over 3,700 peer-reviewed articles have been published using PrimeScript Reverse Transcriptase for a variety of applications, such as RT-PCR gene expression studies, transcriptome analyses, miRNA expression and regulation, molecular evolution, virology, and microbiology.
Overview
- Exceptionally low error rates during reverse transcription and PCR amplification
- Excellent extension capacity even with highly structured template RNAs containing higher-order structures
- Broadly tolerates a wide range of input RNA template concentrations
More Information
Applications
- RT-PCR gene expression studies
- Full-length high-fidelity cDNA synthesis and analysis by endpoint PCR (gel electrophoresis)
- Two-step RT-PCR
- cDNA library construction
Choosing a PrimeScript Kit for endpoint RT-PCR
| If you want to... | Use this... |
| Prepare full-length cDNA using your own primers | PrimeScript Reverse Transcriptase (Cat. # 2680A and 2680B) |
| Prepare full-length cDNA using a kit that includes primers | PrimeScript First Strand cDNA Synthesis Kit (Cat. # 6110A and 6110B) |
| Perform 2-step RT-PCR with a kit that includes RT, primers, and PCR enzyme | PrimeScript RT-PCR Kit (Cat. # RR014A and RR014B) |
| Perform cDNA cloning studies with extremely high fidelity | PrimeScript High Fidelity RT-PCR Kit (Cat. # R022A and R022B) |
| Perform 1-step PCR | PrimeScript One-Step RT-PCR Kit, Ver. 2 (Cat. # RR055A and RR055B) |
Product citations
Cui, X. et al. High expression of osteoglycin decreases the metastatic capability of mouse hepatocarcinoma Hca‐F cells to lymph nodes. Acta Biochim. Biophys. Sin. 40:349–355 (2008).
Matsumoto, T. et al. Role of the aquaporin PIP1 subfamily in the chilling tolerance of rice. Plant Cell Physiol. 50:216–229 (2009).
Qiu, L. et al. Aldose reductase regulates hepatic peroxisome proliferator-activated receptor α phosphorylation and activity to impact lipid homeostasis. J. Biol. Chem. 283:17175–17183 (2008).
Si, D. et al. LplR, a repressor belonging to the TetR family, regulates expression of the L-pantoyl lactone dehydrogenase gene in Rhodococcus erythropolis. Appl. Envir. Microbiol. 78:7923–7930 (2012).
Tanaka, K. et al. Overexpression of HAA1 gene encoding transcriptional activator enhances acetic acid tolerance in Saccharomyces cerevisiae. Appl. Envir. Microbiol. 78:8161–8163 (2012).
Wi, S. J. et al. Synergistic biosynthesis of biphasic ethylene and reactive oxygen species in response to hemibiotrophic Phytophthora parasitica in tobacco plants. Plant Physiol. 159:251–265 (2012).
Wu, F. et al. Endogenous axon guiding chemorepulsant semaphorin-3F inhibits the growth and metastasis of colorectal carcinoma. Clin. Cancer Res. 17:2702–2711 (2011).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
Takara Bio USA, Inc.
United States/Canada: +1.800.662.2566 • Asia Pacific: +1.650.919.7300 • Europe: +33.(0)1.3904.6880 • Japan: +81.(0)77.565.6999
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. © 2024 Takara Bio Inc. All Rights Reserved. All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at takarabio.com.
