I am doing RNA, DNA and protein extraction from the same sample and my protocol recommends this water saturated phenol:chloroform mixture. I am not not very versed with how to go about with the procedure. I will be grateful if I can get some help.
Thanks
Hi there,
All you have to do is to mix 5 vol of liquid phenol with one of chloroform and then mix the mixture with 1 vol water and wait for phase separation (the upper phase is the aqueous phase). Remove the upper phase. Repeat extraction at least twice with fresh water. Store at 4°C with a thin layer of water. Due to volatility, you have to work under a fumehood.
If you don't have liquid phenol then you have to melt it which is very hazardous (heat phenol crystals up to 65°C).
Hello @ Ali Mahmoudpour . I was trying to follow a protocol for simultaneous extraction. It was described by Rajput et al., (2012). They used the method to get DNA, RNA, miRNA and proteins
That's cool.
Dear Ali Mohamoudpour. Your answer was really so informative.
What is differences between common phenol:clorophorm 1:5 and this kind of saturated water solution. We could simply blend the certain vol of each solution to make it..
Dear Ali mahmoudpour
I wholeheartedly admire you for the compressive and of cours informative answers...thanks a million sir.
Dear prof Ali
I'm try to extract rna and DNA from viruses in cell cultur by phenol chloroform but unfortunately nothing extracted or detected by PCR any valid protocol or any precautions to consi please
thanks in advance
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