Q:Why Chloroform in yeast ONPG reaction?

WANGHO at UCBEH.SAN.UC.EDU WANGHO at UCBEH.SAN.UC.EDU
Tue Aug 22 09:43:02 EST 1995


Hi there,

In some protocol about the determination of b-galactosidase activity, the yeast
is washed with Z-buffer and then placed in the Zuffer with SDS and Chloroform.  
I understand the role of SDS.  According to the literature, the chloroform has 
the same role as the SDS does in breaking yeast cells.  My question is why the
chloroform is needed besides the SDS.  Does Chloroform take any other role?

The reason for me to ask this question is that I did the reaction without
chloroform and got much higher value.  By my knowledge, protein can be
denatured by chloroform.  The higher value can be expected.  However, I looked 
up for a lot of protocol and found chloroform in everyone except those methods
using liquid nitrogen or glass beads.

Should I use chloroform or not?

Any suggestion is appreciated.  By the way, I can read through the news group. 
So you can post it in this group or send to me.

Hong Wang
Dept Cell Biology
U Cincinnati Coll. Med.
231 Bethesda Ave (ML521)
CIncinnati, OH 45267-0521



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