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lack of C's cause fading

CRC DNA Sequencing Facility DNASeq at cancer-seqbase.uchicago.edu
Wed Jul 12 12:42:25 EST 2000


Hi,
    Our facility is having a specific problem with templates that have been
generated using bisulfite mutagenesis to convert C residues to T's.  For
templates in which all the C's have been converted to T's, there is fading
that occurs over a span of 200 bases or so which results in complete or
nearly complete loss of signal.  For otherwise identical templates in which
one or a few C's remain, the fading is much less or absent.
    Has anyone else seen this?  Any suggestions on how to modify the
reaction conditions to reduce the fading?

Bill

Sheeba Thomas, Rama Malik & Kari Ensing, Sequencing Technicians
William J. Buikema, Sequencing Director, Ph. D.
Cancer Research Center DNA Sequencing Facility
University of Chicago           Phone: (773)702-1088
920 E. 58th Street            FAX: (773)702-3172
Chicago, IL  60637          E-mail: DNASeq at cancer-seqbase.uchicago.edu
                          Web: http://cancer-seqbase.uchicago.edu/


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