removal of ethanol in PCR product purification - (May/24/2013 )
I am using a qiagen pcr purification kit to purify my pcr produtcs. My problem here is at the last of purification i forgot to centrifuge the column for the second time after adding ethanol to the column so that i can remove all the ethanol that is present. So missing this step i added MQ water to the coulmn and eluted in 30ul.
Now how can i remove this ethanol from the elute ??? After this step i placed elute at 4C.
Please give me some ideas to remove ethanol
thnax in advance
harry
Heat it at 37 degree for 1 hr. I think that will be more than sufficient.
Thanx ashu, but what about he DNA in it?
harry348 on Fri May 24 16:48:50 2013 said:
Thanx ashu, but what about he DNA in it?
What temp. is your body at?
I'm not really sure that this small amount of ethanol (i.e. highly diluted) would preferentially evaporate.
To be completely sure (albeit with the risk of losing some amount of DNA) I would precipitate the eluate with like 75 ul of 100% ethanol, leave overnight at -20 and then centrifuge at 12 000g/10 min/4 degrees and discard supernatant, briefly dry and then dissolve in what amount of water or buffer you want (but note you probably won't see any pelet, but unless it's a really low DNA concentration, it would be there, I use this protocol for desalting PCR products after restriction).
thanks Trof