제품 설명SuperiorScript III Reverse Transcriptase는 M-MLV Reverse Transcriptase (Cat. RT001)의 개량형 제품입니다. 이 제품은 높은 열안정성 (~55°C)과 낮은 RNase H 활성을 보이며, 최대 12kb까지 cDNA를 합성할 수 있습니다. 해당 제품은 Primer (oligo dT, random hexamer) 별도 구매가 필요 합니다.
제품 응용
- Synthesis of first-strand cDNA,
- Array labeling
- cDNA library construction
- 3’ and 5’ RACE, RT-PCR
- Primer extension
제품 구성품
- SuperiorScript III Reverse Transcriptase
- 5X First-Strand buffer
- dNTP Mixture (10 mM each)
- 0.1 M DTT
- Sterile water (RNase free)
제품 품질 관리
- Purity: >99% on SDS-PAGE
- Endonuclease-free
- Exonuclease-free
- RNase-free
- Inhibitor-free
- Satisfactory yield and length of cDNA products
Unit의 정의
One unit is the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insoluble materials using 0.4 mM poly(rA)-oligo(dT) as substrate at 37℃ in 10 min.
Storage buffer
20 mM Tris-Cl (pH7.5), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.01% TritonX-100, 55% glycerol
5X First-Strand Buffer
250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15mM MgCl2
Figure 1. Thermostability of SuperiorScript III on 3 targets
cDNA was synthesized from 500 ng total HeLa RNA for beta-actin 184 bp, GAPDH 395 bp and TFR 738 bp. One-tenth of the cDNA reaction was used for 30 cycles of PCR with nTaq-multi HOT (Cat.# P725HC).
Figure 2. Sensitivity of SuperiorScript III
cDNA was Synthesized from HeLa total RNA with oligo(dT) primer using buffer supplied with each RT and recommended conditions. 10% of cDNA reaction was added to 20 μl PCR reaction containing primers for 500 bp GAPDH gene and 2X TOPsimple™ DyeMIX-HOT (Cat.# P510H), 30 cycle, 1 min/kb.
Figure 3. High sensitivity using SuperiorScript III for RT-qPCR
cDNA was synthesized using SuperiorScript III from 100 ng to 0.1 ng total HeLa RNA for GAPDH 157 bp. cDNA was amplified using TOPreal™ qPCR 2X PreMIX (SYBR Green with low ROX) (Cat.# RT500) on the Bio-Rad CFX-96.
