Abstract
This protocol describes general cloning steps from preparation of both vector and insert DNA to the ligation reaction.
Materials and Reagents
Equipment
Procedure
Recipes
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Hi, Regarding gel purification, any commercial kit of gel purification (i.e. QIAquick Gel Extraction Kit ) can be used. For your second question, not sure what you meant by "any molecular based experiment". But, the amount of each reagent presented in this protocol worked in my hand. Of course, you could adjust the amount of each reagent accordingly if the volume of your reaction assay is different.
Alexander, could you please provide the recipes for the the solutions. In my lab we've been using them for the last year but we can't find the sheet that came with the columns to prepare more buffers. They don't share it on the manufacturer website anymore... Thanks a lot.
Treat vector with alkaline phosphatase before set up the ligation reaction. Please see this link http://www.bio-protocol.com/wenzhang.aspx?id=43&fl3=7 for protocol of using alkaline phosphatase.