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Sequence Analysis of Single Polymorphism in the Factor V Gene - page 3 / 15

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Biol  420                                                                                                                                          Fall 2007

Figure 2: Results of gel electrophoresis for the dye labeled DNA in automated sequencing. The image on the left shows what the gel looks like if the four reactions are run in different lanes, as opposed to the image on the right which shows a gel where all the DNA is run in one lane (Metzenberg).

Since the four dyes fluoresce at different wavelengths, a laser then reads the gel to determine the identity of each band according to the wavelengths at which it fluoresces. The results are then depicted in the form of a chromatogram, which is a diagram of colored peaks that correspond to the nucleotide in that location in the sequence (Russell, 2002).

Figure 3: Results from an automated sequence shown in the form of a chromatogram. The colors represent the four bases: blue is C, green is A, black is G and red is T (Metzenberg).

Pre-lab Questions:

1.

Discuss the difference between Factor V and Factor V leiden? Describe the physiological role of Factor V in clotting.

2.

Describe the process of Sanger dideoxy sequencing.

3.

What is a polymorphism and why is it relevant to this experiment?

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