Base spacing and calling

Karen Heath kheath at hgmp.mrc.ac.uk
Thu Jul 20 14:19:31 EST 2000


Dear Anyone that can help
We are sequencing on an ABI377 and on a 96-lane porous comb.  The last
two gels look great on the gelfile image but when looking at the
sequence on sequencing analysis the first 150-200 is great and then it
looks like a double sequence (ie a bit like a heterozygous mutation) -
some bases are missed, others are doubled and many have N's.  ABI said
it could be the gel mix but we changed all reagents and it still
happened on the second gel - does anyone have any ideas?  Hopefully we
don't have to re-run but can use the software to correct this problem?
If you can help that would be great - we are a core and are running gels
everynight so this is causing a backlog and stress.
Thanks
Karen and Maria
kheath at hgmp.mrc.ac.uk
ramirm04 at doc.mssm.edu


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