blunt end ligation problems

Jeremy C. Ahouse ahouse at hydra.rose.brandeis.edu
Mon Aug 12 15:39:11 EST 1996


Hello group,
   we are having a hard time with blunt end ligation. We are polishing
ends with pfu polymerase. Any suggestions given that? How sensitive are
ligases to salt concentration? I have read about the use of PEG and seen
reference to a protocol that changes the concentration during the ligation
reaction (first high conc favoring insert+vector, then low favoring
circularization). Any thoughts?

   Thanks,

   Jeremy C. Ahouse

-- 
Jeremy C. Ahouse, Biology - Brandeis University
Waltham, MA 01432-1515
617.736-4954 (lab) 617.736-2405 (fax)
ahouse at hydra.rose.brandeis.edu (email)



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