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Dear all,
We are trying to assess preferential incorporation of pro-drugs in DNA or
RNA according to their intratumoral activation.
We are looking for (as usual!) time and cost effective method to separate
DNA and RNA from a same cell preparation.
Data from the literature suggest the use of acid soluble/insoluble method
(cell lysis with diluted PCA, hydrolysis by KOH, reprecipitation of DNA by
concentrated PCA, and that's it) but the method seems so simple and easy
that the authors don't bother in giving details about the different steps.
Attempts to work with this kind of method in our laboratory gave nothing
but crap, so if anybody has a complete and descriptive procedure, it would
be of great help.
Thanks!
Pr. Jacques Catalin, Ph.D.
Laboratoire de Toxicocinetique et Pharmacocinetique
Faculte de Pharmacie
27, Bd Jean Moulin
13385 Marseille cedex 05
France
Phone: (33) (0)4 91 83 55 09
Fax: (33) (0)4 91 83 56 67
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Dear Dr Catalin,
You can refer to "Current Protocols in Molecular Biology" by John
Wiley &
Sons, Inc. There are detailed protocols for DNA and RNA extraction. You
may also consider using commercial kits from Qiagen, Clontech, etc . From
my experience, these kits are easy-to-use, efficient and works fine thus
far.
Lawrence Lee
MPhil Student
Dept of Clinical Oncology
The Chinese University of Hong Kong
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