Question

You are interested in cloning a particular segment of DNA and wish to use a commercial cloning vector with a BamHI cloning site. However, your segment of DNA does not contain the appropriate restriction site. Discussing your problem with a colleague they suggest using PCR to address your problem. How can you use a PCR reaction to facilitate a successful cloning?

Answer 1

Solution:

One of the major objective of cloning is to obtain specific small regions of DNA that contains the gene of interest. For organisms in which their squence is known, amplification using PCR is often the easiest method for getting the gene of interest for cloning. The cloning mthods executed via PCR technique differs from traditional cloning in that the DNA fragment of interest can be amplified by the Polymerase Chain Reaction (PCR) and ligated together, without the use of restriction enzymes. In this context in which your segment of DNA does not contain restriction sites, but you still need it for cloning, definitely you can adopt the PCR technique. One of the major advantage of PCR technique is that it only requires very small amount of DNA for amplification. Being a chain reaction it amplifies the DNA of interest in short span of time.