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religation of vector DNA

Nelson Salazar N.SALAZAR at lshtm.ac.uk
Wed Jul 27 14:41:13 EST 1994


Hello there,
                I am trying to clone two SacII DNA fragments (about 
6.5 and 7.5 kb in size) into pBluescript II separately, and I am not 
getting recombinant plasmid after the transformation. I get many 
colonies but when I screen them, they DO NOT have the insert DNAs 
that I want, that is, the plasmid religates and gives me "false" 
transformants.
 
How could I improve the cloning of my inserts ?? and how could I 
avoid religation of my vector? I think that in this case 
dephosphorylating of the pBluescript SacII cut does not make any 
sense ? Any suggestions will be greatly appretiated.

            Thanks,
            
                        Nelson Salazar

**********************************************************************
Nelson Salazar                      *  E-mail: n.salazar at lshtm.ac.uk
Dept. of Medical Parasitology       *    or  nsalazar at hgmp.mrc.ac.uk
Lon. Sch. Hyg. & Trop. Med.         *  Fax No: 071 636 8739
Keppel St. London WC1E 7HT, UK      *  Tel   : 071 927 2468 
**********************************************************************



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