DNA Science

Slicing and amplifying DNA: restriction digest and PCR.

Using the restiction enyzmes EcoRI and HindIII participants cut Lambda Phage in various lengths and then ran their digest products on an agarose gel (go easy on them...it was their first gel). DNA ladders run with the gel allowed to quantitatively identify each band in our restriction digest. Using NEB's list of restriction sites in the Lambda genome we predicted the cut lengths and were able to identify them all.

The Polymerase Chain Reaction (PCR):

The GFP gene was amplified out of a plasmid using standard PCR, followed by qPCR. In order to illustrate the amplification that takes place during the PCR, the standard reaction was stopped at different cycle numbers and the products run on a gel. For qPCR (quantitative PCR) the fluorescence of a molecule that binds to the product gives a real-time readout of the amplification process.
Doing things the hard way...	... and the easy way.
Looking at amplification the" hard way" is useful to help one appreciate how useful advanced pieces of equipment are, in this case the qPCR machine. Besides the obvious objective of characterizing the amplification of a certain DNA sequence, the idea of this experiment is to get one's hands dirty. Keep in mind this is the first time some students ever used a pipette, which turns an arduous task into something useful.