5 replies to this topic

[Kami23]

Hey all I need to digest my site directed mutagenesis product with dpn1 but want to leave it overnight... will it still work if i do this at 4 degrees or does it HAVE to be 37 degrees for one hour?

[phage434]

Not much will happen at 4C overnight.  You can leave it there, but heat it in the morning for an hour at 37.  You could probably leave it out overnight at room temperature, but I wouldn't do that as a first choice.

[Kami23]

phage434, on Apr 2 2009, 03:31 PM, said:

Not much will happen at 4C overnight.  You can leave it there, but heat it in the morning for an hour at 37.  You could probably leave it out overnight at room temperature, but I wouldn't do that as a first choice.

ok so say i do the digest at 37 deg tonight... would it be ok to freeze it until tomorrow to do my transformation? I mean after the digest just put the lot in the freezer while morning?

Edited by Kami23, 02 April 2009 - 07:06 AM.

[little mouse]

yes, this is what I would do.

[T C]

Hey,
Max I have gone is for two hours.  You can leave the reaction without Dpn I at -20 and do teh Dpn I digestion 2maro..

Best,
TC

Kami23, on Apr 2 2009, 07:53 PM, said:

Hey all I need to digest my site directed mutagenesis product with dpn1 but want to leave it overnight... will it still work if i do this at 4 degrees or does it HAVE to be 37 degrees for one hour?

[Kami23]

T C, on Apr 2 2009, 04:37 PM, said:

Hey,
Max I have gone is for two hours.  You can leave the reaction without Dpn I at -20 and do teh Dpn I digestion 2maro..

Best,
TC

Kami23, on Apr 2 2009, 07:53 PM, said:

Hey all I need to digest my site directed mutagenesis product with dpn1 but want to leave it overnight... will it still work if i do this at 4 degrees or does it HAVE to be 37 degrees for one hour?

damn I already set it going! ah well we shall see what happens