sequencing cosmids

Yuri Trembath tremby at genetics.uiowa.edu
Thu Feb 12 11:25:23 EST 1998


Dear all,

I'm trying to do automated sequencing off a cosmid template and not having
much success (make that ANY success).  If anyone has experience with
primer walking off cosmids and would care to share their advice, I'd love
to hear it.

So far what I've been trying is as follows:  200 ng/uL of cosmid,
linearized with Not I and cleaned up using a Centricon 100 column.  Primer
concentration at 3-10uM.  Primers are designed from cosmid subclones
created via a Sau3A partial digest.  

Please respond to:  tremby at genetics.uiowa.edu

Thanks in advance

Yuri Trembath
The University of Iowa




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