Restrictionenzymelab resultsanddiscussion genesevolutionandecology pdf

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Restriction Enzymes on Bacteriophage Lambda DNA

Results

In determining the known fragment size in base pairs, as shown in Fig. 1, I was surprised

to find that the number of base pairs in the DNA fragment bands for the DNA marker were not

much smaller as they moved further away from the end of the wells at the positive end of the

electrophoresis gel plate. However, they did stay within the limit of 48,502 base pairs as is found

in Bacteriophage lambda DNA.

Figure 1. DNA gel. Labels indicate the content of gel lanes. M-marker (lambda DNA pre-digested with

HindIII), L- uncut Lambda DNA, Pst1- Pst1 Restriction digest of lambda DNA, EcoR1- EcoR1

Restriction digest of lambda DNA, HindIII- HindIII Restriction digest of lambda DNA. A 1% agarose gel

was used and electrophoresis was run at 100V for 30 minutes.

The marker calibration curve, as seen in figure 2 below, was found by plotting the

distance traveled, in millimeters, by each DNA fragment band in the marker lane of the

electrophoresis gel on the x-axis while the log base 10 of the number of base pairs was graphed

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on the y-axis to create a linear curve. The equation generated from the curve in excel displayed a

negative trend, with a negative slope of -.0082, and was used to calculate the number of base

pairings found in the fragmented bands in other lanes in the gel (HindIII, EcoR1, Pst1, and L).

Figure 2. Graph displaying marker calibration curve. This curve was generated using the Marker

(M) data from Table 1 (shown below).

The distance and base pair number data was compiled into Table 1 below for all visible bands in

the electrophoresis gel. Like the calibration curve, the data showed a downward trend. As the

DNA bands moved further away from the wells at the negative end of the plate, the number of

base pairs decreased.

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Restriction Enzymes on Bacteriophage Lambda DNA Results In determining the known fragment size in base pairs, as shown in Fig. 1, I was surprised to find that the number of base pairs in the DNA fragment bands for the DNA marker were not much smaller as they moved further away from the end of the wells at the positive end of the electrophoresis gel plate. However, they did stay within the limit of 48,502 base pairs as is found in Bacteriophage lambda DNA. Figure 1. DNA gel. Labels indicate the content of gel lanes. M-marker (lambda DNA pre-digested with HindIII), L- uncut Lambda DNA, Pst1- Pst1 Restriction digest of lambda DNA, EcoR1- EcoR1 Restriction digest of lambda DNA, HindIII- HindIII Restriction digest of lambda DNA. A 1% agarose gel was used and electrophoresis was run at 100V for 30 minutes. The marker calibration curve, as seen in figure 2 below, was found by plotting the distance traveled, in millimeters, by each DNA fragment band in the marker lane of the electrophoresis gel on the x-axis while the log base 10 of the number of base pairs was graphed 1 on the y-axis to create a linear curve. The equation generated from the curve in excel displayed a negative trend, ...
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