RNA extraction from sols

msho at po-box.mcgill.ca msho at po-box.mcgill.ca
Sun Feb 6 09:16:17 EST 2000


Hi. I am trying to extract RNA, especially mRNA, from microorganisms in
soils. I have tried a method developed by Miskin et al. (using PEG 6000)
and Madsen et al (4 M guanidinium isothiocyanate and ACE buffered-phenol
extraction). So far, I haven't succeeded much since this is my first trial
to extract RNA from soil organisms. I was wondering where I could possibly
make any mistakes that cause degradation of RNA, since I was very careful
not to contaminate my RNA samples from RNase. I soaked all my glasswares,
spatula, and stir bars in 0.1% DEPC water and autoclaved for 45' 30 min at
121 C. Also, all my solutions were autoclaved for at least 30 min. So,
please anyone help me out what part I need to improve. I appreciate any
comments.

Thank you.
---




More information about the Microbio mailing list