RNase protections

STEPHEN A ROLFE S.Rolfe at sheffield.ac.uk
Mon May 9 05:37:09 EST 1994


Dear Arabidopsis folk,

Last week I asked for help on RNase protections on guanidinium
isothiocyanate extracts of plants.  Here are the replies which I
received.  Thanks very much to everyone who replied.

Steve Rolfe
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Check Ambion for doing RNase Protection assay on crude preps.  Their latest
circular, Ambion Technotes vol 1 no.3 has an article on "RNA Analysis with
out RNA Isolation".  I have never tried this so I don't know how well it
works.  If you try it and it works, please let me know!

Carl Garnaat
Pioneer Hi-Bred
GARNAATC at PHIBRED.COM

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We have used an adaptation of Feinberg and Vogelstein 1983 Anal. Biochem.132:
6-13 and Firestein et al. 1987 Anal Biochem. 167:381-386.

The student who worked these methods out for tomato leaf material is Steve
jacobsen. He is no longer in my lab but his E-mail address is:
jacobsens at starbase1.caltech.edu

I suggest you contact him for details.  

Best wishes,
Neil Olszewski

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A method of total RNA extraction, using guanidium
isothiocyanate, and an RNase protection assay using such
total RNA is described in:

Ouellet T, Rutledge RG, Miki B (1992) Members of the
acetohydroxyacid synthase multigene family of Brassica napus
have divergent patterns of expression. The Plant Journal
2(3): 321-330.

          Best wishes,

          George Mourad

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We used total RNA, extracted using guanidium, from a selection of  
Phaseolus tissues (Bennett et al, 1989, Plant Mol Biol, 12, 553-565) 
to study differential expression of the Glutamine synthetase gene 
family by RNase protection

Hope this is useful

Malcolm Bennett
Univ. of Warwick

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