HelixCript™ Thermo Reverse Transcriptase 의 bulk version 입니다.

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HelixCript™ Thermo Reverse Transcriptase 는 M-MLV 역전사 효소(reverse transcriptase, RTase) 유전자에 일부 돌연변이를 도입하여 대장균 세포에서 발현/정제한 열안정성을 증가시킨 재조합 역전사 효소로, 42~55 ℃의 온도에서 주형 RNA로부터 cDNA 합성이 가능하며, 특히 50 ℃에서 최적의 활성을 나타냅니다. HelixCript™ Thermo Reverse Transcriptase 는 높은 processivity와 productivity를 가지고 있어서 RNA 로부터 최대 12kb까지 cDNA를 합성할 수 있습니다.

Application

• Generation of the first-strand cDNA up to 12 kb for libraries or cloning
• Two-step or one-step RT-PCR applications
• Conventional or real-time RT-PCR
• RT-PCR for detection of viral RNA

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 65 ℃

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: 24 months at -20 ℃