double stranded sequencing

Mike Poidinger skspoidn at uk.ac.rdg.susssys1
Thu Jul 16 12:25:12 EST 1992


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Re We've been trying to double-stranded sequence Bluescript inserts ...  
Does anyone have any suggestions?

Rich Buckholz/Mary Martinson
Glaxo Research Institute
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What procedure are you using for the DS denaturing? 
(heating or alkalai, or something else?) Nil sequence 
is probably due to a complete lack of primer binding, 
so it may well be the denaturation which is a problem, 
rather than the annealing per se.

We use alkalai denaturation followed by eppendorf 
spin column desalting, the vital parts of this being
a) length of the spin
b) acid treatment of the glass beads.

Anyway, let me/the net know what exact procedure you use.

Mike Poidinger
Dept of Microbiology
University of Reading

"Use a kit- You Know it Makes Sense"



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