Cleaning PCR Primers

schoenfeld at MSSCC.MED.UTAH.EDU schoenfeld at MSSCC.MED.UTAH.EDU
Mon Dec 12 15:14:52 EST 1994


Jo Caine writes:
>>We have just made some oligos for PCR on a 50pmol scale, cleaved them from 
>>the column, deprotected and dried them down.  When they are redissolved in 
>>water there is visible rubbish in the tube, which does spin down.  I have 
>>tried using the clear supernatant, diluted, in a PCR reaction and get 
>>nothing.  My question is, does anyone know if dirty primers can inhibit the 
>>PCR reaction?  And if so what is the best method (and simplest) for 
>>cleaning them up?>

After resuspending our primers in water, we clean them up with 2 butanol
extractions.  After removing the alcohol (top) layer, measure the O.D. 260
and dilute to your working concentration.

Robert Schoenfeld
Univ. Utah 




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