Plant DNA, Calpain maps?

Oliver S. DePeyer saspeyer at reading.ac.uk
Mon Nov 9 14:01:12 EST 1998


I've got two disparate requests and I'd be very grateful for any advice
anybody could shed on either of them. Please email any suggestions to me
directly (I have trouble accessing the Usenet frequently). I'm at
saspeyer at reading.ac.uk

O.K, problem 1: I have lots and lots of tobacco plants, grown in culture
(i.e in sterile Jars, MSO agar, none of them more than a month old, no
flowers). I suspect that many of them are transgenic. I screened some by
PCR after extracting plant DNA with methods such as Qiaeasy but this isn't
feasible (IMHO) for more than half a dozen plants due to the sheer amount
of time it takes.  What I'd really like to do is something like crushing a
leaf into a PCR tube and run a PCR straight from that - a bit like you can
run PCR from a bacterial colony, or even a mosquito, crushed in a tube. Is
there any protocol like that available for tobacco? 

Problem 2: I have some protein sequences and I need to screen them for
protease cleavage sites, particularly Calpain I & II. What program should
I be using, and which centre could I find it at? (E.g Sanger, NCBI, etc.?)

Looking forward to your advices!

Oliver de Peyer

saspeyer at reading.ac.uk
http://www.rdg.ac.uk/Wolfson

Tel. U.K (0)118 9875123 extn 7889
Fax   "     "   9316671
Pager "  (0)4325 225196

Wolfson Lab (Professor Crabbe's group)
Lab 136
School of Animal & Microbial Sciences (WING)
University of Reading
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PO Box 228
Reading
Berkshire
RG6 6AJ
United Kingdom





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