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ligation for 24 hrs in 4*C?


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3 replies to this topic

#1 kpt.kliper

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Posted 21 October 2009 - 12:36 AM

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

#2 *Jana*

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Posted 21 October 2009 - 03:30 AM

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 16C. Sometimes then the efficience after trafo is higher. So I guess it should matter if you leave it for 24 h at 4C.
Greetings Jana
Beat me if you dare ;) http://krawallziege.mybrute.com

#3 Warren

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Posted 21 October 2009 - 04:50 PM

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..

#4 kpt.kliper

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Posted 22 October 2009 - 03:36 AM

hiya
many thanks people!
kliper




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