Gel Electrophoresis

• Used in vector cloning (to isolate the gene of interest), RFLP (to compare fragments), DNA Sequencing

• Required materials: pBluescript vector plasmid, Restriction Enzymes, Ligase, Bacteria
• Steps
• Cut donor DNA with restriction enzymes, find the targeted DNA fragment through gel electrophoresis 
• Remove section of pBluescript using compatible RE, attach DNA fragment into plasmid at sticky ends, Bind the 2 pieces of DNA using ligase
• Reintroduce pBluescript into the bacterial vector through transformation
• Select bacteria that contains the targeted gene
• Introduce the selected bacteria to a host 

•  Steps
• Cut DNA using RE (blunt)
•  Gel electrophoresis
• Southern blotting (or hybridization) onto a autoradiogram
• Compare sequences of interest 

• Materials: Taq Polymerase, DNA primers, dNTPs, PCR Machine 
• Process: heat --> Cool --> Slight heat = 1 cycle
• The required piece of DNA will be obtained after the 3rd cycle 

• Materials: DNA Polymerase, Radioactive DNA primers, dNTPs, ddNTPs
• All lanes (A T C G) will contain strands of DNA with its complete set of nucleotides