Ligation for two overnights. Is it bad? - (Sep/22/2007 )

Hi there,
I was wondering what would happen if I ligated blunt ended vector and insert two overnights at 14°C. Could it have any disadvantage??

Thank you for your comments.

-clementine-

QUOTE (clementine @ Sep 22 2007, 08:33 AM)

Hi there,
I was wondering what would happen if I ligated by blunt ended vector and insert two overnights at 14°C. Could it have any disadvantage??

Thank you for your comments.

I dont see anything bad can happen unless the possibility of having nuclease contamination. You do have Ca2+ and Mg2+ in the reaction. However, ATP are not very stable at this temperature.

-genehunter-1-

I have left the ligation mixture for 1 week in RT and then transformed it to get positive colonies.

-scolix-