Anyone having problems with PCR Blunt?

Suzanne Suzanne
Mon Jan 20 15:53:20 EST 1997

Invitrogen recently came out with a new plasmid called PCR Blunt. One of
our investigators has just tried it in autosequencing and we are having
dismal results. We sequence with T7 and Reverse primers. The problem with
T7 is that we get nothing, and I do mean nothing. I can't even scare
something up by playing with the raw data.  With reverse we get really
beautiful data up to the insert then bam!,  nothing. We've had the
investigator take some of the plasmid back and try to cut out the insert
just to ensure it's really there and he gets a bright beautiful single
band. I'm inclined to believe this must be a problem with the plasmid. Is
anyone else out there sequencing this plasmid and how are the results? Any
other possibilities I may be missing? Note: These primers work great on
other samples on the same gel.

Suzanne McNeely
DNA Core Facility
smcneely at sunstroke.sdsu.edu

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