PrimeScript RT-PCR Kit
The PrimeScript RT-PCR Kit is a two-step RT-PCR kit featuring PrimeScript RT, an RNase H Minus MMLV reverse transcriptase enzyme that provides robust reverse transcription of virtually any RNA template. This RT-PCR kit is conveniently configured to include all reagents needed for two-step endpoint RT-PCR, including primers, PrimeScript RT, and Takara Ex Taq HS polymerase. PrimeScript RT enables full-length cDNA synthesis from GC-rich and even highly structured RNA templates, while Takara Ex Taq HS allows for efficient, sensitive, and specific cDNA amplification. Reverse transcription can be performed at 42°C, avoiding the risk of RNA degradation, which may occur at higher temperatures. The PrimeScript RT-PCR Kit generates high-yield, full-length cDNA up to 12 kb in length.
- RT-PCR kit includes all reagents needed for two-step endpoint RT-PCR (e.g., where products are analyzed using gel electrophoresis)
- Robust reverse transcription and amplification even from challenging RNA templates
- Complete elongation of complex RNA templates
- High-yield transcription of full-length cDNA up to 12 kb in length
- Minimal false priming and robust amplification due to the combination of PrimeScript RT with Takara Ex Taq HS
- Two-step RT-PCR
|PrimeScript RT (for Two-Step RT-PCR)||25 µl|
|5X PrimeScript Buffer*||200 µl|
|RNase Inhibitor (40 U/µl)||25 µl|
|dNTP Mixture (10 mM each)||150 µl|
|Oligo dT Primer (2.5 µM)||50 µl|
|Random 6 mers (20 µM)||50 µl|
|Takara Ex Taq HS (5 U/µl)||25 µl|
|10X PCR Buffer II*||250 µl|
|Control F-1 Primer** (20 µM)||10 µl|
|Control R-1 Primer*** (20 µM)||10 µl|
|Positive Control RNA (2 x 105 copies/µl)||20 µl|
|RNase-Free dH2O||1 ml|
* 50 reactions of [Reverse transcription (20 µl) → PCR (50 µl)]
** Upstream sense primer for Positive Control RNA
*** Downstream anti-sense primer for Positive Control RNA
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)|
Chen, Z.-Y. et al. Fyn requires HnRNPA2B1 and Sam68 to synergistically regulate apoptosis in pancreatic cancer. Carcinogenesis 32, 1419–26 (2011).
Liu, X. et al. Characterization of the anthranilate degradation pathway in Geobacillus thermodenitrificans NG80-2. Microbiology 156, 589–95 (2010).
Wang, Q., Fang, D.-A., Li, W.-W., Wang, J. & Jiang, H. A novel TCTP gene from the crustacean Eriocheir sinensis: possible role involving metallic Cu2+ stress. Biol. Bull. 221, 290–9 (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.
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