Cleanup of short PCR Product

Emmanuel Skoufos skoufs at minerva.cis.yale.edu
Tue Jun 25 11:20:18 EST 1996


On Tue, 25 Jun 1996, Michael J. McLeish wrote:

> I have a 90 bp PCR fragment that I would like to clean up, sufficiently for 
> direct sequencing.  Most resins seem to have extremely low efficiency for
> fragments of this size.  Any ideas on how to go about this?

I have been succesfully cleaning small PCR fragments (70-100 nt) in the 
following manner:

-Phenol:CHCl3, twice
-Run on a 4% (3:1 Nusieve:SeaKemp ,tms FMC) gel in 1x TBE
-Use the Qiaquick (tm, Qiagen) gel elution columns

I never sequenced *PCR fragments* purified this way (I prefer to clone 
them in TA-type vectors and then sequence); however, in the Qiagen protocol
it is mentioned that DNA cleaned this way is a good sequencing substrate.

> 
> Thanks
> 
> Mike

Emmanuel

(I have no affiliation with the above mentioned companies, just use their 
products:-) )






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