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pcr of total plantcells

suter at de.dbp.MPG.MPIZ-KOELN.VAX suter at de.dbp.MPG.MPIZ-KOELN.VAX
Fri Jul 3 15:31:59 EST 1992


dear colleagues,

at the moment i am doing a lot of RACE and PCR to isolate mRNAs and
genomic DNA from tobacco cells (or tissue). In the past, i did the same
with mammalian cells, and the nice thing was that i could just lyse
these cells in a *special* buffer and do the RT and PCR reaction in one go
(and less than one day).
Tobacco cells are bad for this: they are full of starch and polysaccharides and
other s**t, and also they have a strong cellwall making lysing almost
impossible.
Do any of you have an idea how to simply open these cells so that they are
suitable for PCR right away ? I have tried a simple boiling step already->
but PCR results in fragments <200 bp.

cheers ! clemens
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Clemens Suter-Crazzolara, PhD
Max-Planck-Institut fuer Zuechtungsforschung
Abteilung Genetische Grundlagen der Zuechtungsforschung
Carl-von-Linne Weg 10
5000 Koeln 30
Tel. xx49-221-5062.221           fax. xx49-221-5062.213
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