DEEP WORK

4. PROTOCOL4.1. PreparationEnsure that good-quality DNA preps are available for the receiving vector and any vectors used to retrieve the insert (see Isolation of plasmid DNA from bacteria).  Identify the restriction enzymes needed and their buffer requirements for single and double digests.For PCR product inserts:Add the sequence of any desired restriction site(s) to the 50 end of each primer (..

1. THEORY Molecular cloning is an essential technique to create DNA-based experimental tools for expression in bacterial or mammalian cells. Examples of such DNA constructs include a promoter element fused to a reporter gene or a cDNA sequence under the control of a ubiquitous promoter.  Molecular cloning entails the preparation of the vector and insert DNAs, ligation of the insert into the vect..