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Prepare p:C:I - for RNA extraction (Dec/12/2010 )

Hi,

How can I prepare: phenol: chloroform: iso_amyal alcohol --> 25:24:1, From 5:1 phenol:chloroform?

Thank you
Noor

-noyara-

You can't, if you really insist on the exact amounts. I suppose you could evaporate some chloroform and carefully weigh things to reduce the amount of chloroform before adding isoamyl alcohol. Or you could add a bit more phenol. But that would be stupid. Just add 2% isoamyl alcohol to get to 25:25:1 (well, not exactly, but close enough!). And, just why is this important. Usually phenol/chloroform will work just as well. Major point -- be careful working with this.

-phage434-

phage434 on Sun Dec 12 16:03:54 2010 said:


You can't, if you really insist on the exact amounts. I suppose you could evaporate some chloroform and carefully weigh things to reduce the amount of chloroform before adding isoamyl alcohol. Or you could add a bit more phenol. But that would be stupid. Just add 2% isoamyl alcohol to get to 25:25:1 (well, not exactly, but close enough!). And, just why is this important. Usually phenol/chloroform will work just as well. Major point -- be careful working with this.


Thank you for your suggestion.

-noyara-

noyara on Sun Dec 12 14:13:13 2010 said:


Hi,

How can I prepare: phenol: chloroform: iso_amyal alcohol --> 25:24:1, From 5:1 phenol:chloroform?

Thank you
Noor


add 3.8 chloroform and 0.2 isoamyl alcohol. that should give you the proper ratios.

-mdfenko-

mdfenko on Mon Dec 13 17:13:46 2010 said:


noyara on Sun Dec 12 14:13:13 2010 said:


Hi,

How can I prepare: phenol: chloroform: iso_amyal alcohol --> 25:24:1, From 5:1 phenol:chloroform?

Thank you
Noor


add 3.8 chloroform and 0.2 isoamyl alcohol. that should give you the proper ratios.


Hmm, I c, Thank you very much,

By the way, I have phenol crystal in the lab, but as I read in internet, not recommended to use crystal form, Is there any way to prepare stock solution from crystal form?
Many thanks in advance

-noyara-

see the cold spring harbor protocols page on phenol.

-mdfenko-