본 제품은 HelixCript™ Thermo Reverse Transcriptase의 bulk 공급 품목입니다. 

대량 구매를 원하시면 sale@nanohelix.net으로 문의주시기 바랍니다. 

HelixCript™ Thermo Reverse Transcriptase는 RNase H-negative M-MLV 역전사효소(reverse transcriptase, RTase) 변형체입니다. 열에 안정적인 이 RTase는 42 ~ 55℃의 온도 범위에서 RNA 주형으로부터 cDNA를 합성할 수 있으며, 50℃에서 가장 높은 활성을 나타냅니다. 본 제품의 우수한 역전사 효율은 template RNA에서 최대 12kb까지 cDNA를 합성할 수 있습니다.


Application

• cDNA library 제작 또는 클로닝을 위한 최대 12 kb의 1^st strand cDNA 생성
• two-step / one-step RT-PCR
• RT-PCR / real-time RT-PCR
• 바이러스 RNA 검출

 

Figure 1. Comparison of activity of HelixCript™ Thermo Reverse Transcriptase with a competitor by RT-PCR.

Total RNAs were extracted from human blood. The first-strand cDNAs were synthesized using total 700 ng RNA, oligo-d(T) primer, 200 U HelixCript™ Thermo Reverse Transcriptase at 50 ℃ or 200 U Promega’s M-MLV RTase (Promega, USA) at 42 ℃. 10 % of cDNA reaction was used with a primer set corresponding to fragments of human dynein gene and 2.5 U HelixAmp™ HyperSense-Taq DNA Polymerase in 50 μl PCR reaction.

Enzyme activities: RNA-dependent DNA polymerase. No RNase H activity

 

Recommended reaction temperature: 42 to 55 ℃

 

Storage buffer: 20 mM Tris-HCl (pH 7.5), 100 mM KCl, 0.1 mM EDTA-NaOH (pH 8.0), 1 mM DTT, stabilizers, 50 % glycerol

 

Purity: ≥ 90 % (SDS-PAGE)

 

DNase contamination test: Not detectable (Incubation with 200 U enzyme and pUC19 plasmid at 37 ℃, 1 hr)

 

RNase contamination test: Not detectable (Incubation with 200 U enzyme and human total RNA at 37 ℃, 1 hr)

 

Activity test in cDNA synthesis, RT-PCR and qRT-PCR: Correspond to reference (Human RNA target)

 

Stability: 12 months at -20 ℃