Storage of BigDye sequencing reaction samples

Mart Speek smart at kbfi.ee
Tue Mar 9 18:21:30 EST 1999


Hey "BigDye" SEQUENCERS!

Recently I had a serious problem  when using BigDye kit based
sequencing reactions and running  ABI 310 machine.
Got no sequence!
(details were posted in bionet.molbio.methds -
http://www.bio.net/hypermail/METHDS-REAGNTS/)

Now it turned out to be related to the storage of
TSR-treated DNA samples at -20 degrees Centigrade.

When identical samples were stored at -20 C and +4 C
refrigerators for one week,
only +4 samples showed good sequences (reads up to 600 without N's),
while -20 C samples gave no peaks, NO SEQUENCES!!!
Similarly, fresh and good DNA samples gave nothing
after storage at -20C for 5-10 days.

I have no idea what happened to these
reactions at -20 C.

Would be interested to hear whether anyone else has
encountered similar phenomenon.

Mart (ESTONIA)




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