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Ligation with PLAFR - (Jun/09/2004 )

Please help
I am trying to ligate fragments of DNA that have been double digested with Bgl II and Hind III into the vector PLAFR3.
I am making my cells competent by washing in Calcium chloride and using freeze thaw to transform them.
can anyone offer sugesstions of why this is not working mad.gif

-alshia-

Hi alshia,

Many things in your experiment could have gone wrong, such as ligation, preparation of competent cells, and transformation. Before others could answer your question, please provide some details:

What type of your DNA is, PCR product or DNA cut from from a vector?
How did you prepare your vector?

-pcrman-