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
follow:

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 
genome.

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 
cDNA.

C Cho Yue
ccy at po.cwru.edu
-- 



More information about the Methods mailing list