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Products >  Real_Time_qPCR_Kits >  RT-qPCR_Reagents >  PrimeScript_RT_Reagent_Kit_with_gDNA_Eraser_for__Real_Time_PCR

Eliminate Genomic DNA Contamination: PrimeScript RT Reagent Kit with gDNA Eraser

Genomic DNA contamination can be a significant problem during real-time RT-PCR because co-amplification of genomic DNA and cDNA can lead to erroneous results. With the PrimeScript RT Reagent Kit with gDNA Eraser, genomic DNA contamination is eliminated. This reverse transcription kit enables fast cDNA synthesis for real-time RT-PCR (qPCR) and includes a rapid step that eliminates genomic DNA (gDNA) contamination without loss of input RNA.

The PrimeScript RT Reagent Kit with gDNA Eraser includes PrimeScript Reverse Transcriptase, a modified, RNase H Minus recombinant MMLV (Moloney Murine Leukemia Virus) enzyme which allows fast cDNA synthesis due to its exceptionally strong strand displacement activity. PrimeScript RTase is robust, versatile and can be used to prepare cDNA from almost any RNA template, including GC-rich templates and RNAs with significant secondary structure.  Because of the excellent extension capacity of PrimeScript, cDNA synthesis reactions can be performed at lower temperatures (i.e., 42°C), decreasing the risk of RNA degradation.

In the first step, genomic DNA contamination is eliminated by treating the samples with gDNA Eraser (an optimized, potent DNA degradation enzyme) for 2 minutes at 42°C. The subsequent reverse transcription reaction includes a component that completely inhibits DNA degradation activity, and reverse transcription is complete in just 15 minutes. The final result is high quality cDNA with no trace of genomic DNA contamination. This kit is designed for two-step RT-PCR and should be used with Takara Bio qPCR premixes 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).

First introduced in Japan, PrimeScript RTase and the kits containing this enzyme are now available worldwide. Over 3,700 peer-reviewed articles have been published in which PrimeScript Reverse Transcriptase is applied towards the study of gene expression, gene discovery, transcriptome analysis and miRNA expression regulation.   

  At-A-Glance   Documents   Images & Data   Resources

Features

  • Genomic DNA contamination elimination in only 2 minutes
  • Fast cDNA synthesis - reverse transcription in just 15 minutes
  • Suitable for use with routine and challenging RNA templates alike
  • Full length cDNA synthesis up to 12 kb

Applications

  • Elimination of genomic DNA contamination
  • cDNA synthesis for real-time RT-PCR (qPCR)
  • Two-step real-time RT-PCR

RR047A (100 reactions (20 μL))

gDNA Eraser100 μL
5X gDNA Eraser Buffer*1200 μL
PrimeScript RT Enzyme Mix I*2200 μL
5X PrimeScript Buffer 2 (for Real Time)*3400 μL
RT Primer Mix*4400 μL
RNase Free dH2O1 mL x 2
EASY Dilution (for Real Time PCR)*51 mL

*1: Because 5X gDNA Eraser Buffer is needed for the subsequent reverse-transcription reaction, please be sure to perform the genomic DNA elimination reaction.

*2: Contains RNase inhibitor.

*3: Contains dNTP mixture.

*4: Contains Oligo dT Primer and Random 6 mers.

*5: Use when preparing serial dilutions of total RNA and cDNA. EASY Dilution makes it possible to obtain a precise dilution down to very low concentrations of nucleic acid.  Moreover, this solution will not affect reverse transcriptase or PCR polymerase enzyme activity. The diluted template solution may be used directly for reverse transcription and PCR reactions.

EASY Dilution (for Real Time PCR) may also be purchased separately (Cat. #9160).

Note:Use this kit in combination with Takara Bio’ s Real-Time PCR kits. Compatibility with products from other manufacturers has not yet been verified.

RR047B (400 reactions)

This kit is comprised of 4 x RR047A.

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)

References

1. Fukui, K. et al. (2013) Mol. Plant 6:88-99.

2. Nakajima, A. et al. (2012) Appl. Envir. Microbiol. 78:6251-6261.

3. Nemoto, T. & Shibasaki, T. (2012) Am. J. Physiol. Endocrinol. Metab. 302: E781-E787.

4. Qin, P. et al. (2013) Plant Cell. Physiol. 54:138-154.

5. Sasaki, N. et al. (2013) Physiol. Mol. Plant Pathol. In Press. http://dx.doi.org/10.1016/j.pmpp.2013.08.002

6. Sato, Y. et al. (2012) J. Biol. Chem. 287:23236-23245.

7. Wang, Q. et al. (2012) J. Biol. Chem. 287:6159-6168.

8. Xiang, J.-J. et al. (2012) Plant Physiol. 159:1488-1500.

9. Yang Y. et al. (2012) Appl. Envir. Microbiol. 78:5845-5854.  

10. Zhou, M.-L. et al. (2012) Glycobiol. 22:1775-1785.

11. Zhou, Z. et al. (2012) J. Anim. Sci. 90:3752-3761.




 
 
Products
Cat. # Product Contents Size Price Units Select
RR047A PrimeScript™ RT Reagent Kit with gDNA Eraser (Perfect Real Time) 100 Rxns $425.00
RR047B PrimeScript™ RT Reagent Kit with gDNA Eraser (Perfect Real Time) 400 Rxns $1,446.00
 

 

Recommended Products
Cat. # Product Contents Size Price License 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
RR037A PrimeScript™ RT Reagent Kit (Perfect Real Time) 200 Rxns $282.00  
RR037B PrimeScript™ RT Reagent Kit (Perfect Real Time) 800 Rxns $901.00  
RR390A Premix Ex Taq™ (Probe qPCR) 200 Rxns $233.00 License Statements
RR390B Premix Ex Taq™ (Probe qPCR) 400 Rxns $463.00 License Statements
RR036A PrimeScript™ RT Master Mix (Perfect Real Time) 200 Rxns $425.00  
RR036B PrimeScript™ RT Master Mix (Perfect Real Time) 800 Rxns $1,365.00  
 

 

 
 

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