Generating "good" blunt ends

Nino nivil at gengenp.rug.ac.be
Mon Nov 20 04:34:27 EST 1995


I cloned some cosmids randomly in M13 to generate DNA sequencing, we shear
these cosmids by sonication and lately by nebulizing the DNA, afterwards we
made a DNA ends repair by using T4 DNA Pol. and Klenow enzyme and we separate
according to MW. by agarose gel. (~1100 bp.)
Our control is with lambda DNA digested with AluI enz.
My problem is the low efficiently of cloning after T4 and Klenow treatment.
Any suggestions are WELLCOME.




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