产品编号： K1632    

产品名称：  RevertAid H Minus First Strand cDNA Synthesis Kit/逆转录（RT）试剂盒   

供应商：  MBI

规格： 100 react

目录价： 2800  

相关信息： COMPONENTS OF THE KIT

RevertAid™ H Minus First Strand cDNA Synthesis Kit 20 rxns

#K1631

100 rxns

#K1632

RevertAid™ H Minus M-MuLV Reverse Transcriptase (200 u*/μl) 25 μl 120 μl

RiboLock™ RNase Inhibitor (20 u**/μl) 25 μl 120 μl

5X Reaction Buffer

250 mM Tris-HCl (pH 8.3), 250 mM KCl, 20 mM MgCl2, 50 mM DTT 150 μl 500 μl

10mM dNTP Mix 50 μl 250 μl

Oligo(dT)18 Primer

100 μM, 0.5 μg/μl (15 A260 u/ml) 25 μl 120 μl

Random Hexamer Primer

100 μM, 0.2 μg/μl (6 A260 u/ml) 25 μl 120 μl

Forward GAPDH Primer, 10 μM

5’ – CAAGGTCATCCATGACAACTTTG – 3’ 20 μl 20 μl

Reverse GAPDH Primer, 10 μM

5’ – GTCCACCACCCTGTTGCTGTAG - 3’ 20 μl 20 μl

Control GAPDH RNA

1.3 kb 3’-poly(A) tailed RNA transcript, 0.05 μg/μl 20 μl 20 μl

Water, nuclease-free 2x1.25 ml 2x1.25 ml

* One unit of RevertAid™ H Minus M-MuLV RT incorporates 1 nmol of dTMP into a polynucleotide fraction

(adsorbed on DE-81) in 10 min at 37°C.

** One unit of RiboLock™ RNase Inhibitor inhibits the activity of 5 ng RNase A by 50%.

STORAGE

All components of the kit should be stored at -20°C. Keep control RNA at -70°C for longer

storage.

DESCRIPTION

The RevertAid™ H Minus First Strand cDNA Synthesis Kit is a complete system for efficient

synthesis of first strand cDNA from mRNA or total RNA templates. The kit uses RevertAid™

H Minus M-MuLV Reverse Transcriptase, which has a point mutation that completely

eliminates RNase H activity. Therefore, degradation of RNA does not occur during first strand

cDNA synthesis, resulting in higher yields of full-length cDNA from long templates (up to 13kb)

compared to other reverse transcriptases. The enzyme maintains activity over a wide

temperature range (42-55°C) which makes it an ideal tool for reverse transcription of RNAs

having a high degree of secondary structure.

The recombinant RiboLock™ RNase Inhibitor, supplied with the kit, effectively protects RNA

from degradation at temperatures up to 55°C.

The kit is supplied with both oligo(dT)18 and random hexamer primers. Random hexamer

primers bind non-specifically and are used to synthesize cDNA from all RNAs in total RNA

population. The oligo(dT)18 primer selectively anneals to the 3’-end of poly(A) RNA,

synthesizing cDNA only from poly(A) tailed mRNA. Gene-specific primer may also be used

with the kit to prime synthesis from a specified sequence.

First strand cDNA synthesized with this system can be directly used as a template in PCR or

real-time PCR. It is also ideal for second strand cDNA synthesis or linear RNA amplification.

Radioactively and non-radioactively labeled nucleotides can be incorporated into first strand

cDNA for use as a probe in hybridization experiments, including microarrays.

 

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注：该产品未在中华人民共和国食品药品监督管理部门申请医疗器械注册和备案，不可用于临床诊断或治疗等相关用途