Jose de las Heras
josenet at tiscali.co.uk
Thu May 4 14:21:41 EST 2006
By PCR *and* digest? why both?
you can find a restriction site close to one end in your insert, and either
with that alone or in combination with another in the vector you should
generate a pattern that will tell you in which orientation the fragment is.
But it's easier by PCR alone, if you can wait to get the primers, one from
the vector and another from the insert, as others explained. It's the
"newsnet customer" <customer at newsnet.com> wrote in message
news:ZQk6g.22547$vy1.3229 at news-server.bigpond.net.au...
I have transformed GFP gene into a vector.
Then transformed the vector into competant E.coli cells.
I have many colonies.
How to check if colonies have my gene in the CORRECT orientation by PCR and
restriction digest? I think the only way is to sequence the region in both
Any comments appreciated.
More information about the Methods