IUBio

Poor resolution on 377 to 300bps only in odd lanes

McNeely, Suzanne suzanne.mcneely at invitrogen.com
Thu May 31 18:44:18 EST 2001


We run a small core facility using a 377 sequencer. Lately we have begun to
notice a pattern developing where our samples in even lanes do very well but
the samples in the odd lanes are very poor. It appears that the first 300
bps or so the peaks are broad and run over each other, after that the data
seems normal. We tested this by running the same sample in both the even and
odd lanes and the evens look good and the odds are bad. We run a 4.5% LR gel
overnight(10hrs). We heat the samples for two minutes prior to loading and
place on ice. We load the odds first, prerun for 2 minutes and then load the
evens. This is our standard procedure and has been for the past two years.
Has anyone seen anything like this? Any suggestions as to what might be
causing this phenomenon?

Thanks,

Suzanne

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