the effect of tRNA on cDNA cloning ?

Shahram Mori smori at nmsu.edu
Wed Dec 13 21:05:22 EST 1995


Seongwook Kwon (swkwon at email.unc.edu) wrote:
: Hi,

: I am thinking of using tRNA to help out precipitating my cDNAs.  These 
: cDNAs are to be used for a library construction.

: 	Does anyone know the effect of tRNA presence on 1) conversion of ss 
: cDNA into ds cDNA (I will use Klenow), 2) ligation of insert DNA into 
: lambda vector 3) packaging of vector-insert into lambda particles ?


You can do two things if tRNA's bother you;

1) Use 10mg/ml glycogen (add 5ul to your DNA) and then add NaOAC and 95% 
COld ETOH

Or

2) Add a little Rnase to the resuspended pellet.
Cheers
--
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Shahram Mori
Program in Molecular Biology
Department of Chemistry and Biochemistry Box 3C
New Mexico State University
Las Cruces NM
88003
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