Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Cloning

please help!!! with transformation - (Nov/05/2009 )

Pages: Previous 1 2 

phage434 on Nov 9 2009, 12:07 AM said:

pIAlac is Erm resistant to 300 ug/ml according to the paper Perez-Arellano02, Structural features of the lac promoter affecting gus expression in Lactobacillus casei. The E. coli origin of the vector is a p15A origin, which is low copy number, and may mean you are having difficulty preparing plasmid DNA. There is more than enough information in that paper to allow full sequencing, which would be the first thing I would do if I were serious about working with the plasmid.

Try 1:10 ratio of vector:insert. I suceeded ligation of 1.3 kb insert in 10 kb vector

Pages: Previous 1 2