Mon Feb 1 15:37:00 EST 1993

We have put insertions in several as yet unidentified chromosomal
genes of E.coli, all regulated by a common regulator. We would like
to identify them by sequencing out from the end of the insertion,
taking advantage of the fact that 44 % of E. coli is now sequenced.
Our insertions so far are made with lambdaplacmu.

Has anybody tried this with this, or any other insertion? I am told
that the left end of mu is more difficult to handle than the right.
WE propose to make a specific primer and a universal primer,
amplify by PCR and then sequence. Any information, experiences,
useful primer sequences, and ideas for other more convenient
insertion elements would be gratefully received.

With thanks,
EBNewman  Biology Dept. Concordia University. Montreal, Canada.

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