IUBio

Poor resolution on 377 to 300bps only in odd lanes

Dr N.I.Leaves nleaves at hgmp.mrc.ac.uk
Thu Jun 7 07:03:58 EST 2001


Interesting. Try loading the evens first... and then the odds. I can only
tihnk its something to do with your pre-running which would be the only
difference between the otherwise identical samples.

Nick

end 
************************************************** 
Dr N I Leaves
Mouse Sequencing
MRC HGMP Resource Centre 
Hinxton 
Cambridge CB10 1SB 
tel: 01223 494557 (office) or 01223 494541 (lab)
email: nleaves at hgmp.mrc.ac.uk
**************************************************

On 1 Jun 2001, McNeely, Suzanne wrote:

> 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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