T vector clone sequencing problem help!!!

Roland Hubner roland.hubner at ua.ac.be
Mon Jul 29 09:43:14 EST 2002


> Sequence result shows the DNA sequence
> is not my sequence ,but size and including prime
> sequence are the same with mine.who can tell me any
> tip about this .Thank for any help!!
> 
> Bright Yeh 

Hello,

 can you not further increase your annealing temperature to obtain
specific PCR product? 

 Alternatively, use "touchdown PCR" or new primer(s) might
be necessary...

 Q: How do you 'hotstart' your reactions? Is your sequence GC-rich?

Good luck,
 Roland



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