ABI seq'g and DH10B cells

David N. Smith david.smith at cbr.for.csiro.au
Mon Jan 23 21:17:21 EST 1995


Has anyone successfully sequenced a clone propogated in DH10B (MAX
Efficiency DH10B Competent Cells from GIBCO BRL), using cycle sequencing on
the ABI 373 automated sequencing system?

Does anyone know if this E. coli strain allows recombination between
plasmids that contain tandem repeats?

I am having a hell of a time trying to get reasonable sequence from a
library containing microsatellites that was produced in these cells. I have
had successful results using DH5alpha, unfortunately this particular library
was made in DH10B. I have tried to eliminate all other possible causes of
seq'g failures on the ABI 373. I suppose I am really clutching at straws in
order to explain what is happening in the reactions. I have yet to 1) try to
transform DH5alpha cells with the same library, and 2)try to sequence
something from DH10B that doesn't contain a microsatellite (when it works it
works fine!).

Thanks,
David.




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