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.
