You mean you want to linearise the construct by cutting it with a RE outside
the expression cassette before transformation??? Is that what you mean. I
don't see the purpose of cutting a vector after ligating vector and fragment to
clone a PCR fragment efficiently. It does not make much sense.
Jay
-----Original Message-----
From: [email protected]
[mailto:[email protected]] On Behalf Of Nasser Mahna
Sent: Tuesday, December 22, 2009 12:23 PM
To: [email protected]
Subject: Cloning trick
Dear All,
I would to ask if there is anybody mastered in working with restriction and
digestion.
I want to clone a pcr fragment inside a binary vector. To do it efficiently, I
want to use a restriction enzyme after ligating vector and fragment, which cuts
the original uncut vector before transformation.
what method do you offer?
Best regards,
Nasser.
_______________________________________________
Methods mailing list
[email protected]
http://www.bio.net/biomail/listinfo/methods
This email message may contain legally privileged and/or confidential
information. If you are not the intended recipient(s), or the employee or
agent responsible for the delivery of this message to the intended
recipient(s), you are hereby notified that any disclosure, copying,
distribution, or use of this email message is prohibited. If you have received
this message in error, please notify the sender immediately by e-mail and
delete this email message from your computer. Thank you.
_______________________________________________
Methods mailing list
[email protected]
http://www.bio.net/biomail/listinfo/methods
