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Products >  Real_Time_qPCR_Kits >  RNA_Extraction_for_qPCR >  CellAmp_Whole_Transcriptome_Amplification_Kit_for_RT_PCR_Real_Time

Cells to cDNA: CellAmp Whole Transcriptome Amplification Kit for RT-PCR (Real Time) Ver. 2

The CellAmp Whole Transcriptome Amplification Kit (Real Time), Ver. 2 enables one to go from cells to cDNA and then use the generated cDNA as a template for real-time PCR.  When using conventional techniques for working with trace amounts of nucleic acids, loss of product during purification steps is a frequent problem. However, this cells to cDNA kit ensures good product yield because loss-inducing purification steps for RNA and cDNA are not required.

The procedure begins with cell lysis, followed by a reverse transcription reaction with the dT adapter primer (RT dT Primer 2) to synthesize cDNA from mRNA. Next, terminal deoxynucleotidyl transferase (TdT) is used to add a dA tail to the synthesized cDNA. The resulting 3' dA tail containing cDNA product is used as direct template for PCR, resulting in fewer required amplification cycles.

In addition to moving from cells to cDNA, this product can be used to amplify cDNA from trace amounts of RNA.

The CellAmp Whole Transcriptome Amplification Kit (Real Time), Ver. 2 uses an additional exonuclease I treatment step, which helps suppress nonspecific primer-derived amplification and increases yields of low expression level genes.

At-A-Glance Documents Images & Data Resources

Features

Applications

  • Cells to cDNA with minimal purification
  • Real-time RT-PCR (RT-qPCR)
  • cDNA amplification from trace amounts of RNA

Components

Lysis Buffer (4×)150 μL 
Recombinant RNase Inhibitor (40 U/μL)30 μL
RT dT Primer 212 μL
dNTP Mixture (2.5 mM each)12 μL
MgCl2 (22.5 mM)36 μL
RT Enzyme Mix 2*136 μl
Exonuclease I (5 U/μL)72 μL
TdT Buffer (5×)144 μL
dATP (90 mM)24 μL
TdT Enzyme Mix*254 µL
PCR Primer Mix 2300 µL
RNase Free dH2O1 mL

*1: Contains PrimeScript Reverse Transcriptase and RNase Inhibitor

*2: Contains Terminal Deoxynucleotidyl Transferase and RNase H

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
3734 CellAmp™ Whole Transcriptome Amplification Kit (Real Time), Ver. 2 100 Rxns $434.00
 

 

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