Restriction Enzyme Lab Report

Purpose: Restriction enzymes cut DNA at a certain palindromic sequence. Three samples of lamda DNA set up to be cut with restriction enzymes PstI, EcoRI, or HindDIII. There were also two more samples, one of these samples was not mixed with any restriction enzyme and the other was a marker, which used an enzyme which creates fragments with a known number of base pairs used to create a standard curve. All five samples were put through agarose gel electrophoresis in order to estimate the amount of base pairs of the fragments in the non marker samples using the standard curve created with the marker sample. It was hypothesized that if three samples of DNA are mixed with different restriction enzymes and 1 sample is not mixed with a restriction enzyme and agarose gel electrophoresis is performed on all 4 samples, then the patterns shown in the gel will be different and the sample with with no restriction enzyme will have only one band. This was hypothesized because each of the three restriction enzymes cut DNA at a different palindromic sequence which are located at different parts in the DNA, therefore the fragments created by each enzyme should have different numbers of base pairs which would cause them to travel different distances in the agarose
The number of base pairs was already known for these fragments, but the distance had to be measured using figure 1. A standard curve was used to predict the number of base pairs in the fragments of the 4 other lanes, the estimated values can be found in table 1. From these values in table 1 and the drawing in figure 1 it can be seen that the patterns of the three restriction enzyme lanes were significantly different and that the lane with no restriction enzyme only had one