jehane, on 02 September 2010 - 04:04 PM, said:
bob1, on 02 September 2010 - 03:56 PM, said:
The same as you would for any other chloroform extraction - add 1 volume of chloroform to the sample, shake or vortex vigourously for 30 s, then centrifuge to separate layers...
without need for phenol:chloroform step at first?
Yes, just add chloroform.
The purpose of phenol is to denature proteins. Since proteins aren't a problem here, there is no need for phenol:chloroform mixture.
However if you don't have chloroform and only have phenol-chloroform, you can use phenol-chloroform mixture instead.
May your PCR products be long, your protocols short and your boss on holiday