Subject: Neurospora RNA for RNase protection

From: [email-masked] (rlab davis)
Date: Sun, 12 Mar 1995 19:14:03

Is anyone using a particular RNA isolation protocol for Neurospora which 
yields total RNA suitable for RNase protection?  I seem to be having trouble 
with DNA contamination using techniques published in the FGN.

Ideally, I would like a technique which is quick, easy and doesn't involve 
isolating mRNA from total.  Is this wishful thinking?

If you have any suggestions, please email me.  Thanks in advance.

Martin Hoyt 
Dept. of Molecular Biology and Biochemistry, Univ. Calif. Irvine
[email-masked]

---

From: [email-masked] (Matthew Sachs)
Date: 13 Mar 1995 12:45:29 -0800

>Dept. of Molecular Biology and Biochemistry, Univ. Calif. Irvine

1.  Lower pH (5-6) in aqueous buffers for RNA extraction is reported to
reduce the recovery of contaminating DNA.

2.  A simple but not quantitative method to separate DNA (and small RNA)
from larger RNA, such as mRNA, is on the basis of their differential
solubilities in lithium chloride.

3. RNase-free DNase can be used to treat total RNA samples.

See:

Wallace, D. M. (1987). Large- and small-scale phenol extractions. Meth.
Enzymol. 152, 33-41.

Wallace, D. M. (1987). Precipitation of nucleic acids. Meth. Enzymol. 152,
41-48.

-----------------------------------------------------------
Matthew Sachs
Department of Chemistry, Biochemistry and Molecular Biology
Oregon Graduate Institute of Science and Technology
20000 NW Walker Road
P.O. Box 91000
Portland, OR  97291-1000
503 690-1487 Phone
503 690-1464 Fax
[email-masked]