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  • ‹ Back to PrimeScript cDNA synthesis kits
  • PrimeScript IV first-strand cDNA synthesis mix
  • PrimeScript two-step kit
  • PrimeScript high-fidelity two-step kit
  • PrimeScript II 1st Strand cDNA Synthesis Kit
  • PrimeScript 1st strand cDNA Synthesis Kit
  • PrimeScript one-step kit
  • PrimeScript Double Strand cDNA Synthesis Kit
Home › Products › mRNA and cDNA synthesis › cDNA synthesis kits › PrimeScript cDNA synthesis kits › PrimeScript two-step kit

cDNA synthesis kits

  • PrimeScript cDNA synthesis kits
    • PrimeScript IV first-strand cDNA synthesis mix
    • PrimeScript two-step kit
    • PrimeScript high-fidelity two-step kit
    • PrimeScript 1st strand cDNA Synthesis Kit
    • PrimeScript one-step kit
    • PrimeScript Double Strand cDNA Synthesis Kit
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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.

Cat. # Product Size Price License Quantity Details
RR014B PrimeScript™ RT-PCR Kit 200 Rxns USD $1224.00

This kit for two-step RT-PCR is designed to provide excellent extendibility and efficient amplification. It uses PrimeScript reverse transcriptase and Takara Ex Taq HS polymerase. This kit generates RT-PCR products efficiently at standard reverse transcription temperature (42℃), even when RNA templates contain higher-order structures. It is not necessary to perform reactions at a high temperature when there is a risk of RNA decomposition. This kit includes all reagents necessary for the reverse transcription of RNA to cDNA, and for cDNA amplification using PCR. Cat. # RR014B contains 4 of Cat. # RR014A. Please refer to Cat. # RR014A for complete product documentation and resources.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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RR014B: PrimeScript RT-PCR Kit

RR014B: PrimeScript RT-PCR Kit
RR014A PrimeScript™ RT-PCR Kit 50 Rxns USD $362.00

This kit for two-step RT-PCR is designed to provide excellent extendibility and efficient amplification. It uses PrimeScript reverse transcriptase and Takara Ex Taq HS polymerase. This kit generates RT-PCR products efficiently at standard reverse transcription temperature (42℃), even when RNA templates contain higher-order structures. It is not necessary to perform reactions at a high temperature when there is a risk of RNA decomposition. This kit includes all reagents necessary for the reverse transcription of RNA to cDNA, and for cDNA amplification using PCR.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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RT-PCR using various amounts of RNA template

RT-PCR using various amounts of RNA template

RT-PCR using various amounts of RNA template. To illustrate the high sensitivity of the PrimeScript™ RT-PCR Kit in detecting mRNAs in very low quantities of total RNA template, GAPDH mRNA was used as a target in various amounts of HL60 cell total RNA (0.1 pg to 1 ng). The RNA was reverse transcribed using the oligo dT primer. The kit protocol was followed for RT and PCR.

Protocol:
Target mRNATotal RNA
GAPDH (428 nt)HL60 cell
RT reaction: 0.1 pg –> 1 ng total RNA/20 µl reaction
PCR: 2 µl RT products/50 µl reaction
PCR conditions:
94°C –> 1 min.
(40 cycles)
94°C –> 30 sec.
55°C –> 30 sec.
72°C –> 1 min.
Results:
Detection of the GAPDH target mRNA was possible using as little as 1 pg of total RNA.

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Amplifying various lengths of target gene mRNAs by RT-PCR

Amplifying various lengths of target gene mRNAs by RT-PCR

Amplifying various lengths of target gene mRNAs by RT-PCR. Using human heart total RNA or HL60 cell total RNA as a template, a variety of sequence lengths from two target gene mRNAs (dystrophin and transferrin receptor) were reverse transcribed and amplified by PCR using the reagents in the PrimeScript™ RT-PCR Kit.

 
Protocol:
Target mRNATotal RNA
DystrophinHuman Heart
Transferrin receptor (TFR)HL60 cell
 
Reverse transcription: 1 µg total RNA/20-µl reaction
PCR: 2 µl RT products/50-µl reaction*
----
*Corresponds to 100 ng total RNA/50-µl PCR reaction
 
PCR conditions:
0.5~6kb8~12 kb
94°C –>1 min94°C –> 1 min
(30 cycles)(30 cycles)
94°C –> 30 sec.98°C –> 10 sec
55°C –> 30 sec.68°C –> 8 or 15 min
72°C –> 1 min./kb
Results
Excellent yields were obtained from all reactions (see figure), confirming efficient extension and amplification of sequences between 0.5 kb and 12 kb in length.

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Principle of PrimeScript RT-PCR Kit

Principle of PrimeScript RT-PCR Kit

Principle of PrimeScript RT-PCR Kit.

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RR014A: PrimeScript RT-PCR Kit

RR014A: PrimeScript RT-PCR Kit

Overview

  • 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

More Information

Applications

  • Two-step RT-PCR

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

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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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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