Description:
The cDNA Synthesis Kit is designed for preparation of
full-length first strand cDNA from RNA templates. The kit relies
on a cloned enzyme, Moloney murine leukemia virus reverse
transcriptase (M-MuLV RT). M-MuLV RT synthesises first strand
cDNA at sites determined by the type of primer used:1. Random
hexamer primer: at non-specific points along an RNA template. In
this case, all RNA in a population are templates for cDNA
synthesis.
2. Oligo(dT)18: at the 3*-end of
poly(A)+ mRNA. In this case, only mRNA with 3*-poly(A) tails are
templates for cDNA synthesis.
3. Sequence specific primer: at a primer-binding site.
First strand cDNA synthesized with this system can be used as
a template in the polymerase chain reaction (PCR). As the
reaction conditions of cDNA synthesis and PCR are compatible,
the cDNA reaction mixture can be added directly to a PCR
mixture. The kit is optimized for maximum yield of full length
cDNA. The addition of a ribonuclease inhibitor lowers the risk
of mRNA degradation during the reaction.
The first stand cDNA synthesized may also be used as a
template for second strand synthesis. Radio-labeled DNA can be
added as a probe in hybridization experiments.
As most users do not need to do control first strand cDNA
synthesis, this kit do not include control RNA.
Notes:
- Multiple freezing and thawing of RNA should be
avoided. Thaw and keep control RNA on ice.
- It is recommended that the first strand cDNA
synthesis is carried out under conditions where RNase
contamination has been eliminated. Pipette tips and tubes should
be treated with 0.1% DEPC.
- Wearing gloves is highly recommended.
This product is distributed
for laboratory research only.
Caution: Not for diagnostic use . |
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