DNase treatment of RNA sample

Toshiharu Ishizuka tishizuk at AQUARIUS.BEKKOAME.OR.JP
Tue Mar 14 06:58:24 EST 1995


Dear bionetters

        I would like to thank all people who responded my problem. Most
people recommended me the pilot experiments to determine which component
contains RNase. I did the experiments and found that the buffer for DNase
treatment contains RNase. Thus, I would like to ask you. 
        How can I prepare Tris-based DNase buffer without RNase ? I heard
that diethylpyrocarbonate reacts with Tris, and it is difficult to
inactivate RNase by autoclaving. 

The composition of our DNase I buffer is 
10mM Tris-Cl, pH8.3
50mM KCl
1.5mM MgCl2
10U human placental RNase inhibitor (TOYOBO)
10U RNase-free DNase I

I appreciate if anybody teach me.

Sincerely yours,

Dr. Toshiharu Ishizuka
Department of Biochemistry, Chiba University, School of Medicine
1-8-1, Inohana, Chuo-ku, Chiba, 260 JAPAN
FAX +81-43-226-2041
E-mail
QFH03407 at niftyserve.or.jp
tishizuk at aquarius.bekkoame.or.jp




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