Description

This kit is designed to perform cDNA amplification directly from small amounts of cell. The amplified cDNA can be applied for real-time PCR as a template. In general, there is a problem in loss of nucleic acids during purification process when dealing with small amounts of nucleic acid. However, cDNA can be amplified efficiently without purifying RNA or cDNA by using this kit.
In this kit, perform cell lysis at first, followed by synthesize cDNA from mRNA by reverse transcription using dT adapter primer (RT dT Primer). Next, add dA tail by TdT to synthesized cDNA, which will be used as a template for low cycle PCR and amplify the cDNA.
This kit is useful to directly amplify cDNA from cells, and also to amplify cDNA from small amount of RNA.

This product is not available in the U.S.

Components

(for 50 reactions)

1.	Lysis Buffer	350 µl
2.	RNase Inhibitor (40 U/µl)	25 µl
3.	RT dT Primer	20 µl
4.	dNTP Mixture (2.5 mM each)	20 µl
5.	RT Enzyme Mix*1	50 µl
6.	TdT Buffer	300 µl
7.	dATP (100 mM)	30 µl
8.	TdT Enzyme Mix*2	25 µl
9.	PCR Primer Mix	600 µl×2
10.	RNase Free dH2O	1 ml×2

*1 Contains PrimeScript™ Reverse Transcriptase and RNase Inhibitor
*2 Contains Terminal Deoxynucleotidyl Transferase and RNase H

Storage

-20°C

Reagents and instruments not supplied in this kit

• PCR reagent
  TaKaRa Ex Taq™ Hot Start Version (Cat. # RR006A/B)
• 1.5 ml microtube
• 0.2 ml microtube
• Micropipette and tips
• Thermal Cycler
  TaKaRa PCR Thermal Cycler Dice™ (Cat. #TP600/TP650) etc.