strange cloning result

Wolfgang Schechinger Wolfgang.Schechinger at med.uni-tuebingen.de
Mon Mar 27 16:51:45 EST 2000


Hi all, 

recently, I performed a simple cloning experiment (sticky 
HindIII-XbaI insert into pCDNA3). 
As directed in the textbooks, I had an control ligation without the 
insert. This control gave me about 300 colonies as "background". The 
sample containing the insert yielded 100 colonies only of which I 
prepped 24. Finally 15 (60%) contained the insert though I didn't 
expect to get any positive. Is there any reasonable explanation for 
this except statistics? Was it an insert overdose killing most bugs 
due to too long insert concatenations? (Concentrations were estimated 
on gel, insert to host ratio was 5)

Wolfgang
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Dr. Wolfgang Schechinger, Dept. of Pathobiochemistry
University of Tuebingen, Germany
email: wolfgang.schechinger at med.uni-tuebingen.de 
wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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