Request for assistance with extraction of chloroplast DNA from moss Ceratodon

Matt Shelton mshelton at GDI.NET
Wed Nov 6 11:59:51 EST 1996


Lake Brantley High School is participating in the joint Ukraine-NASA CUE
mission.  Students in dual-enrollment Principles of Biotechnology have
selected mini-research projects which they are pursuing, many under the
mentorship of scientists via the Internet.  Lisa Harris is interested in
comparing the chloroplast DNA of Brassica rapa (Wisconsin fast plants)
and the moss Ceratodon purpureus.  Her hypothesis is that there is a
different restriction fragment pattern between these two plants.  Lisa
is well-versed in gel electrophoresis, restriction digests, etc.  As we
do not have a thermocycler, she has decided to proceed with chloroplast
isolation.  Dr. Christine Chase has provided a copy of the protocol
developed by Hughes, Eaton, and Thompson at Furman University.  We have
the following question in regard to this protocol:
	1.  The homogenization medium is NaCl, Tris, EDTA, BSA, and 
            MeSH.  What is MeSH?  Can it be purchased from
Sigma             Chemical Co?
	2.  Proteins and residual genomic DNA is precipitated with KAc.
	    What is KAc?  Does Sigma carry it?
	3.  Need the e-mail and/or FAX for company selling Spectramesh.    	4. 
Above protocol suggests use of restriction 	    endonucleases: PstI and
AvaI.  We also have AvaII, BglI, 	    	    HaeII, HincII, EcoRI,
HindIII, and BamHI.  Are these worth
	    trying?

Any guidance you can provide will really be appreciated.



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