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cytotrap library construction problem

Frederik Börnke ricky_boernke at gmx.net
Fri Jan 21 08:10:44 EST 2000


Hi all,

we are desperately trying to construct a cDNA library from tobacco
for Stratagene's CytoTrap THS. However, although we yield plenty of
cDNA (1.5 - 2 µg) ligation efficiency into the precut pMyr vector
is very poor. We tried several ratios between vector and insert without
any positiv effect. The controls for ligation and transformation 
are quiete persuasive. Thus, it seems that there is something wrong
with our inserts, like inefficient linker ligation or else. Has
anybody encountered trouble like this? Would an additional round
of linker ligation help? Any hints will be appreciated.
BTW, if anybody has a library prepared from plants and would be
willing to share such a library, we would be very pleased.

TIA
Daniel and Ricky

*******************************************************************

Frederik Boernke
Research Group of  Molecular Plant Physiology
Institute for Plant Genetics and Crop Plant Research (IPK)
Corrensstr. 3
06466 Gatersleben
Tel.  039482 -5 321
Fax. 039482 -5 515
http://www.ipk-gatersleben.de




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