Liyana Ismail wrote:
Hi Catherine,
I'm tempted to reply to this one since we're doing something pretty similar.
Yup, this should be a workable protocol in the general case. The
problem here is that Catherine only has 10-15bp of overlap between the
two original products, as compared to your 319bp. That will make the
joining reaction much more difficult, since you can't do it at PCR
temperatures.
To be honest, with that small a degree of overlap, Catherine might be
better placed incorporating appropriate restriction sites into her
primers and then cutting/ligating. It depends on how critical the exact
sequence of the joining region is and whether she has the leeway to
design in a RE site.
Peter
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