HelixCript™ 1st-Strand cDNA Synthesis Kit는 total RNA 및 poly(A) RNA로부터 1^st strand cDNA를 효율적으로 합성하는데 필요한 모든 구성요소를 포함한 제품입니다. 본 제품에 포함되어있는 HelixCript™ Thermo Reverse Transcriptase는 M-MLV RTase의 RNaseH-negative variants로 열에 안정(thermostable)하며, 활성 온도는 42 ~ 55℃ 입니다. 높은 온도에서 RNA의 2차 구조 형성이 저해되고, RTase의 중합 활성이 증가하여, cDNA 합성 활성이 증가됩니다. 본 제품에 포함된 RNase inhibitor template RNA가 Reaction Mix 내에서 분해되지 않도록 하여 cDNA 합성 효율을 높여줍니다. 합성된 1^st strand cDNA는 별도의 추가 처리 과정 없이 PCR이나 real-time PCR의 주형으로 직접 사용 가능합니다.

 

 

Figure 1. RT-PCR using HelixCript™ 1st-Strand cDNA Synthesis Kit and random hexamer or oligo-dT as primers for reverse transcription. 1st-strand cDNA synthesis reactions were performed using the 300 ng total RNA isolated from human blood. 10% of each 1st-strand cDNA products were used in PCR reactions with primer sets corresponding to fragment of human dynein gene and 1.25 units HelixAmp™ Xtender DNA polymerase in 50 ul PCR reaction.

Application 

• Generation of first strand cDNA for subsequent PCR or Real-time PCR 
• cDNA library construction

 

 

Contents 

• 50 pmole/㎕ oligo d(T20) primer
• 50 pmole/㎕ random hexamers 
• dNTP mix (each 10mM)
• 5x RT reaction buffer
• 0.1M DTT
• RNase inhibitor 
• HelixCript™ Thermo Reverse Transcriptase (200unit/㎕)
• DEPC water

 

 

Quality Control

HelixCript™ 1st-Strand cDNA Synthesis Kit was functionally tested for RT-PCR amplification on different sized products (0.5 ~ 5 kb)

with 500 ng of total RNA. 500 bp RT-PCR product was obtained from 100 pg of total blood RNA using human β-actin primers.