Primer stocks that fail to PCR over time

Robert Preston rapr at med.pitt.edu
Thu Nov 24 15:27:39 EST 1994


In article <CzqnrD.4Jz at ncifcrf.gov>, pnh at fcsparc6.ncifcrf.gov (Paul N
Hengen) wrote:

> them in their -20 C freezer. If you've had trouble with this problem and feel
> like sharing your experience to be written in an upcoming TIBS column, please
> post or e-mail. Thanks.

Paul, et al., I'm discovering loss of signal with 5'-fluorescent-tagged primers
stored at 10pMol/ul in TE for six months then used for PCR under identical
conditions. Not clear whether the label is going bad or the amplification not
working or both.  Signals are down two to three or more orders of magnitude
depending on the primer set, these being Genethon/CEPH human microsatellite
markers that behaved very well in the past.  I have a couple hundred sets,
not all appear to have gone bad, although I haven't examined the phenom. yet
for more than a dozen sets. Oh, temp was -20C for storage, in a non-frost-free.

-- 
Robert Preston                                    rapr at med.pitt.edu
Highland Park 40deg28'N 79deg55'W               Pittsburgh PA 15206
U. Pittsburgh Sch. Med. S792 Scaife Hall        Pittsburgh PA 15261
   vox 412-648-9573  fax 412-648-1916 (W)  vox 412-361-8905 (H)



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