Geneclean and cDNA synthesis

Jorge L. Sepulveda Jorges at bcm.tmc.edu
Wed Dec 18 17:45:34 EST 1991


In article <009534BC.CAECCBA0 at m44.unm.edu> kim at m44.unm.edu writes:
> I was wondering if anyone had an opinion of the idea of using Geneclean 
or some
> homebrew of the same to concentrate and change buffer systems in a more
> conventional cDNA synthesis system.  This would afford a significant time
> savings as well in any cDNA synthesis system.


I have tried to purify single-stranded DNA with the glass beads with no 
success. Also, I have had variable recoveries with double-stranded DNA 
(10-50%, as measured by radiactivity), especially with smaller fragments.

On the other hand, a 15 minutes ethanol precipitation  at -70!C, with 
carrier (tRNA, glycogen, etc) always yields more than 90% recovery. This 
takes about the time used by glass bead isolation, with half the hassle.

I wouldn't trust my precious cDNA to the glass beads :-)

Jorge Sepulveda

Jorge L. Sepulveda
Dept. Pathology .......................... (713)-798-7330
Baylor College of Medicine
1 Baylor Plaza, Houston, TX 77030



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