Plasmid sequencing

"bruce pearson IFR bruce.pearson at BBSRC.AC.UK
Mon Jun 4 08:50:24 EST 2001


Dear Knut,
          I would imagine your best approach would be to shotgun clone 4
base cutter fragments into a pUC18 or 19 and sequence a number of clones.
This would give you a good starting point for making primers to PCR regions
and to  primer walk. The number of initial clones you would need would
obviously depend on the size of the plasmids. The advantage however of the
shotgun phase is the rapid rate at which get the sequence. Shift to the
primer walking when you start duplicating sequence. 

Cheers Bruce

Bruce M Pearson
BBSRC Institute of Food Research
Norwich Research Park
Colney Lane
Norwich UK
Tel 01603 255196
email Bruce.Pearson at BBSRC.AC.UK 

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