baffling pBluescript stuff...

Ed ed at bio02.bio.uottawa.ca
Wed Nov 27 14:28:11 EST 1996


hey all,

herer's the deal :

- had some positive cDNA clones in Lambda ZapII

- tried invivo excision of the pBluescrit phagemid - seems to have worked 
because I had tons of colonies on LB + AMP plates

- now I try sequencing two of the clones with vector primers but I get 
nothing...not a single thing !!!

the frustrating thing is that I can't even hypothesize some massive 
deletion or anything because I have cut out the inserts with enzymes that 
are on opposite ends of the MCS. The primers I'm using for sequencing 
also seem fine because two other clones from the same library screening 
have sequenced beautifully. It's just that the two clones I can't 
sequence are the ones that I would want most dearly for they are the ones 
with longest inserts.

I could kill something/someone...

please enlighten me...


Ed Taboada
Molecular Genetics
Dept of Biology
University of Ottawa
Ottawa, Canada



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