Can PCR be used to nonspecifically applify cDNA's?
Cheung C. Yue
ccy at po.CWRU.Edu
Mon Feb 10 10:32:38 EST 1992
In a previous article, IO00865 at MAINE.MAINE.EDU () says:
>Is it possible to amplify cDNA's before cloning using nonspecific PCR?
> (stuff deleted)
It is definitely feasible. Most protcols that I have seen which deals
with cDNA do not use ligation of adapters to double-stranded cDNA
perhaps due to the problem of loss of 5' sequences. Most involve
tailing reactions, but one recent one use RNA ligase. References
PNAS 85:8998 '88. Classic paper on the RACE protocol (Rapid Amplifi-
cation of C-DNA Ends).
Science 243:217 '89. A nice application of anchored-PCR (RACE) to
study of T-cell receptors.
Methods in Mol and Cell Biol 2:17 '90. Describes a nice approach to
limit the size of cDNA for more even amplification using anchored-PCR.
PNAS 88:9623 and 9628 '91. Two similar papers illustrating the
creative use of general amplification of cDNA's in dissecting the
PCR Methods and Applications 1:43 '91. RNA ligase instead of tailing
to get the 5' end of specific cDNA. I have not thought enought about
whether this approach could work on a heterogeneous population of
C Cho Yue
ccy at po.cwru.edu
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