3 replies to this topic

[deespike]

hi all
im working on blunt end ligation .i have searched this forum for some tips on this ligation but im confused with the temperature usage for the reaction.what is the optimum temoerature for performing blunt end liagtion?

thanks in advance

[WHR]

deespike, on Jun 1 2009, 02:39 AM, said:

hi all
im working on blunt end ligation .i have searched this forum for some tips on this ligation but im confused with the temperature usage for the reaction.what is the optimum temoerature for performing blunt end liagtion?

thanks in advance

T4 ligase is most active at 37 degree C.  I usually perform ligation reaction at RT for convenience.

Edited by WHR, 01 June 2009 - 06:48 AM.

[madrius1]

People sometimes do it at 16 C O/N. Really depends on your needs/time!

[T C]

Hey,

Read this too:

http://www.protocol-...re for ligation

Best,
TC

deespike, on Jun 1 2009, 04:09 PM, said:

hi all
im working on blunt end ligation .i have searched this forum for some tips on this ligation but im confused with the temperature usage for the reaction.what is the optimum temoerature for performing blunt end liagtion?

thanks in advance