About ligation

KHUANG at CHEMVX.TAMU.EDU KHUANG at CHEMVX.TAMU.EDU
Fri Apr 19 18:15:36 EST 1996


Hi, Netters,
I have a few silly question to ask you.
(1) About ligation temperature. 
Any enzyme (?) should have its best activity temperature. Some protocol 
said we can do ligation at 4, 16, 22-25 degree, It is said T4 DNA ligase 
has a few best activity temperature, right?
(2) About the size of insert
It is difficult to do ligation for longer insert. For pBluescript, how 
longest insert can be ligated (I heard of 21 kb)? Do you think it is
possible to find (or creat) new ligase for blunt, long insert ligation et al?
(3) What is the secret in the 30 min ligation kit? New ligase or inducer 
exist there?
Thanks 

Huang



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