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Products >  PCR_Products >  Real_Time_qPCR_Kits >  RT-qPCR_Reagents >  PrimeScript_RT_Reagent_Kit

Generate cDNA for real-time PCR: PrimeScript RT Reagent Kit (Perfect Real Time)

When synthesizing cDNA for real-time PCR, the PrimeScript RT reagent Kit (Perfect Real Time) allows maximal flexibility during reaction assembly. This reverse transcription reagent kit facilitates cDNA synthesis prior to analysis by real-time PCR. Designed for two-step real-time RT-PCR (also called qRT-PCR or RT-qPCR), the PrimeScript RT Reagent Kit (Perfect Real Time) contains all of the components needed for reverse transcription, including PrimeScript RTase, RNase Inhibitor, Random 6mers, Oligo dT Primer, dNTP Mixture and reaction buffer. Because the kit's primers and RTase are provided as separate components, there is greater flexibility in reaction assembly. If a master mix is preferred, consider using PrimeScript RT Master Mix (Perfect Real Time. Use this kit in combination with one of Takara Bio's qPCR premix products, such as SYBR Premix Ex Taq II (Tli RNase H Plus) (Cat.# RR820A), SYBR Premix Ex Taq (Tli RNase H Plus) (Cat.# RR420A), or Premix Ex Taq (Probe qPCR) (Cat.# RR390A), for SYBR Green or TaqMan probe-mediated detection.

The PrimeScript RT Reagent Kit (Perfect Real Time) includes PrimeScript Reverse Transcriptase for strong strand displacement activity and efficient synthesis of cDNA for real-time PCR. PrimeScript RTase is a modified, recombinant MMLV (Moloney Murine Leukemia Virus) reverse transcriptase and is verified to be RNase H Minus. PrimeScript RTase is also robust, versatile and can be used for reverse transcription of virtually any RNA template, including templates that have a high degree of secondary structure or are GC-rich.  Because of the excellent extension capability of PrimeScript, cDNA synthesis reactions can be performed at 42°C, minimizing the risk of RNA degradation that often occurs at higher temperatures.

At-A-Glance Documents Images & Data Resources

Features

  • Allows fast, efficient synthesis of cDNA for real time PCR. This kit is designed for two step real-time RT-PCR.
  • Includes Random 6 mers and Oligo dT Primer for use as reverse transcription primers. The reaction can be performed using one or a mixture of these two primer types; alternately, gene specific primers may be used.
  • Includes EASY Dilution (for Real Time PCR) when generating reference standards for a qPCR standard curve. This reagent facilitates higher accuracy at lower standard concentrations.

Applications

  • Synthesis of cDNA for real-time PCR

Components

(for 200 reactions) There is a partial modification from Lot AK1101 as to the Kit Components.

5X PrimeScript Buffer (for Real Time)1 400 µL
PrimeScript RT Enzyme Mix I 100 µL
Oligo dT Primer50 µM100 µL
Random 6 mers100 µM400 µL
RNase Free dH2O 1 mL
EASY Dilution (for Real Time PCR)2 1 mL

1:contains dNTP Mixture and Mg2+.
2:

It can be used to prepare the dilution series of total RNA or cDNA, which is required for establishing the standard curve.

In contrast to dilution with water or TE, EASY Dilution facilitates preparation of low-concentration standards. EASY Dilution does not inhibit enzyme activity of reverse transcriptase or PCR polymerase.

EASY Dilution is also available separately: EASY Dilution (for Real Time PCR) (Cat.# 9160)

Note: Use this kit with qPCR premixes from Takara Bio. Compatibility with products from other manufacturers has not been verified.

Note

A standard curve must be generated for quantitation by real-time PCR. Dilution of total RNA or cDNA is necessary in order to include low concentration samples, which are important for a viable standard curve with an appropriately wide range. However, dilution with water or TE can narrow the range of the curve because nucleic acids are unstable at low concentrations in the presence of dH2O or TE. Use EASY Dilution (for Real Time PCR) provided in this kit for generating accurate reference standards.

Storage

–20°C

Choosing a PrimeScript Kit for Real-Time RT-PCR

If you want to....

Use this....

Make cDNA using a master mix that includes RTase and primers

PrimeScript RT Master Mix (Perfect Real Time) (Cat. # RR036A and RR036B)

Make cDNA using a kit that provides RTase and primers as separate components

PrimeScript RT Reagent Kit (Perfect Real Time) (Cat. # RR037A and RR037B)

Avoid genomic DNA contamination during cDNA synthesis and RT-qPCR

PrimeScript RT Reagent Kit with gDNA Eraser (Perfect Real Time) (Cat. # RR047A and RR047B)

Perform 1-step real-time RT-PCR with SYBR Green

One-Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time) (Cat. # RR086A and RR086B)

Prepare cDNA for RT-qPCR directly from fewer than 1,000 cells

CellAmp Whole Transcriptome Amplification Kit (Real Time), Ver. 2 (Cat. # 3734)




 
 
Products
Cat. # Product Package Size Price # of Units Select
RR037A PrimeScript™ RT Reagent Kit (Perfect Real Time) 200 Rxns $282.00
RR037B PrimeScript™ RT Reagent Kit (Perfect Real Time) 800 Rxns $901.00
 

 

Recommended Products
Cat. # Product Package Size Price License # of Units Select
9160 EASY Dilution (for Real Time PCR) 8 x 1 mL $128.00  
RR420A SYBR® Premix Ex Taq™ (Tli RNase H Plus) 200 Rxns $290.00 License Statements
RR420B SYBR® Premix Ex Taq™ (Tli RNase H Plus) 400 Rxns $566.00 License Statements
RR820A SYBR® Premix Ex Taq™ II (Tli RNase H Plus) 200 Rxns $290.00 License Statements
RR820B SYBR® Premix Ex Taq™ II (Tli RNase H Plus) 400 Rxns $566.00 License Statements
RR390A Premix Ex Taq™ (Probe qPCR) 200 Rxns $233.00 License Statements
RR390B Premix Ex Taq™ (Probe qPCR) 400 Rxns $463.00 License Statements
 

 


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