Subcloning - (May/01/2001 )
I would appreciate it if you could give me any ideas you may have on the subject as soon as possible, preferably by tuesday, may 2. My question is on subcloning. In a class, we subcloned DNA into a bacterial plasmid. We expected the insertion of the DNA to be in random orientation. However, only 1 person managed to subclone it with the 5' end of the gene near the forward primer of the plasmid. the remaining 20 students cloned it with the 3' end near the forward primer of the plasmid. Any suggestions on why this may have occurred? Thank you very much for your time.
The product may be toxic to the cell
that orientation may have been favourable in the ligation, or it was just luck!