Unstable DNA spacers

Brian Halligan halligan at post.its.mcw.edu
Tue Sep 28 09:45:00 EST 1993


We are interested in introducing 'spacer' DNA into a construct so we can
test the effects of distance from the end of the molecule to an internal
site. We have tried introducing linkers of 10 bp in length, but it appears
that these tandem linkers are unstable, ie. clones that had 5 linkers for
a 50 bp spacer, when restreaked produced clones with 3, 2, and 1 insert,
but none with 5 inserts.  For this experiment, we would like to have a
distribution of distances from the internal site to the end and keep a
consistant end so that cutting and labeling could be done in a standard
fashion for all the clones.  The size range we would like would be inserts
from 10 to 200 bp.  We have thought of introducing random DNA fragments,
but would rather avoid having to sequence all of the clones or having to
use multiple clones of the same size to avoid effects due to the random
sequence. 

Any suggestions?


Brian D. Halligan			halligan at post.its.mcw.edu
Assistant Professor			414-257-8413 (voice)
Department of Microbiology		414-257-8427 (FAX)
Medical College of Wisconsin
-- 
Brian D. Halligan			halligan at post.its.mcw.edu
Assistant Professor			414-257-8413 (voice)
Department of Microbiology		414-257-8427 (FAX)
Medical College of Wisconsin



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