3 replies to this topic

[kpt.kliper]

Hiya folks,
I have a question - can I leave my ligation reaction (pgemt) in 4*C for ca 24 hrs? I know they recommend an overnight reaction, but can anything go wrong if I leave it for  24 hrs?
best regards
kliper

[*Jana*]

kpt.kliper, on Oct 21 2009, 10:36 AM, said:

Hiya folks,
I have a question - can I leave my ligation reaction (pgemt) in 4*C for ca 24 hrs? I know they recommend an overnight reaction, but can anything go wrong if I leave it for  24 hrs?
best regards
kliper

I often do a ligation for more than 24h at 16°C. Sometimes then the efficience after trafo is higher. So I guess it should matter if you leave it for 24 h at 4°C.
Greetings Jana

[Warren]

kpt.kliper, on Oct 21 2009, 04:36 AM, said:

Hiya folks,
I have a question - can I leave my ligation reaction (pgemt) in 4*C for ca 24 hrs? I know they recommend an overnight reaction, but can anything go wrong if I leave it for  24 hrs?
best regards
kliper

from experience I am sure you can leave it at least a few days at 4 degrees, I wouldn't suggest it for long term storage though.....I have seen claims that too long a ligation time can cause problems in a buffer with PEG (something about creating giant oligimers and subsequent interference with transformation) but I haven't run into that problem.  24 hours should be about the same as 16 hours ("overnight") at 4 degrees.

Warren..

[kpt.kliper]

hiya
many thanks people!
kliper