신제품 Induro™ Reverse Transcriptase는 열 안정성 및 억제제 내성이 증가된 역 전사 효소입니다. 기존 제품 보다 빠른 시간 안에 long cDNA를 얻을 수 있으며 긴 전사체, 2차 구조를 가진 RNA, inhibitors가 있는 RNA 샘플에도 사용할 수 있습니다.
- Highly processive RT enables fast reactions with high yields of long cDNA
- Strong inhibitor tolerance enables robust cDNA synthesis performance
- Supports direct RNA sequencing and long read cDNA sequencing workflows
- Increased thermostability for robust cDNA synthesis at high temperatures
- Experience comparable fidelity to retroviral RTs
Induro™ Reverse Transcriptase
Induro Reverse Transcriptase is a group II intron-encoded RT that exhibits high processivity, increased thermostability, and increased tolerance of inhibitors in the synthesis of cDNA from RNA. It is an ideal enzyme for challenging cDNA synthesis from long transcripts, RNAs with strong secondary structures, and RNA samples with inhibitors. With improved 5′ sequencing coverage of long transcripts, Induro Reverse Transcriptase can enable RNA-seq applications such as direct RNA and long read cDNA sequencing workflows.
Figure 1: Induro Reverse Transcriptase exhibits high processivity, permitting rapid cDNA synthesis
Induro Reverse Transcriptase can synthesize a full-length 12 kb cDNA product within 5 min. at 55°C. In vitro transcribed poly(A) RNA templates (1 kb, 4 kb, 8 kb or 12 kb) were used to investigate full-length cDNA synthesis. After first-strand cDNA synthesis, RNA was hydrolyzed immediately by NaOH. Subsequently, an aliquot of the cDNA products was used to make full-length ds cDNA in the presence of a 5´ specific primer. Equal volume of ds cDNA was analyzed on an agarose gel.
Figure 2: Thermostability of Induro Reverse Transcriptase allows robust synthesis at higher reaction temperatures
Induro Reverse Transcriptase can make a full-length 12 kb cDNA product in 10 min. using incubation temperatures of 45°C to 60°C. In vitro transcribed poly(A) RNA templates (1 kb, 4 kb, 8 kb or 12 kb) were used to investigate full-length cDNA synthesis. After first strand cDNA synthesis, RNA was hydrolyzed immediately by NaOH. Subsequently, an aliquot of the cDNA products was used to make full-length ds cDNA in the presence of a 5´ specific primer. Equal volume of ds cDNA was analyzed on an agarose gel.
Figure 3: Induro Reverse Transcriptase generates highest yields of long cDNA
Induro Reverse Transcriptase generates the highest product yields for cDNA ≥ 8 kb. RNA templates were in vitro transcribed poly(A) RNA (1 kb, 4 kb, 8 kb, 10 kb or 12 kb). After first strand cDNA synthesis, RNA was degraded and the second strand cDNA synthesis was performed in the presence of a 5´ specific primer.
NEB 사에서 판매되는 역 전사 효소(RT)에는 레트로바이러스 (M-MuLV, MMLV)로 인코딩된 RTs와 인트론 인코딩 (group II intron-encoded) 된 RT 두 가지 유형이 있습니다. 신제품 Induro™ Reverse Transcriptase는 Retroviral RTs와 비교하여 higher thermostability, faster processivity를 갖습니다.
|
|
Intron-encoded RT |
Retroviral RT Products |
||
|
Product |
Induro™ Reverse Transcriptase (NEB #M0681) |
LunaScript RT SuperMix Kit (NEB #E3010) LunaScript RT SuperMix (NEB #M3010) |
LunaScript RT Master Mix Kit (NEB #E3025) – Primer-free version |
ProtoScript® II First Strand cDNA Synthesis Kit (NEB #E6560) |
|
Applications |
ㆍLong cDNA synthesis ㆍImpure RNA samples ㆍRNA-seq applications (e.g., direct RNA sequencing, long-read cDNA sequencing, tRNA seq |
ㆍ2-step RT-qPCR detection ㆍRNA-seq workflow |
ㆍRNA-seq workflow ㆍ2-step RT-qPCR detection ㆍGeneral cDNA synthesis ㆍ2-step RT-PCR detection |
ㆍGeneral cDNA synthesis ㆍRNA-seq workflow ㆍ2-step RT-PCR detection |
|
Format |
Standalone Reagents |
Supermix |
Supermix |
Reaction mix, enzyme mix, primer |
|
Reaction Temperature |
Up to 60°C (55°C optimal) |
Up to 65°C (55°C optimal) |
Up to 65°C (55°C optimal) |
Up to 48°C (42°C optimal) |
|
RNA Input |
up to 1 µg |
up to 1 µg |
up to 1 µg |
up to 1 µg |
There are mainly two types of Reverse Transcriptases (RT): retrovirus-encoded RTs and intron-encoded RTs. Both RT types share some conserved domains and synthesize complementary DNA from an RNA template. The most common and well-characterized RTs are derived from retroviruses such as Moloney Murine Leukemia Virus (M-MuLV, MMLV), such as ProtoScript II Reverse Transcriptase.
In contrast, Induro Reverse Transcriptase is a group II intron-encoded RT that exhibits higher thermostability, faster processivity, and better tolerance of reaction components when compared to retroviral RTs. Induro Reverse Transcriptase is useful for cDNA synthesis from long transcripts, RNAs with strong secondary structures, and RNA samples with inhibitors.
The recommended incubation time for Induro Reverse Transcriptase is 10 minutes at 55°C. Longer incubation times up to 30 minutes at 55°C may improve the yield of long cDNA products.
