Name: ____________________________

BIO 181: Lab Quiz #7 (10 pts) – Gel Electrophoresis

Questions 1-4, please refer to the following: DRD4 is an autosomal gene that encodes a receptor
for the neurotransmitter dopamine. The length of this protein differs among individuals due to
variation in the number of repeats of a 48 base pair (bp) insertion in the third exon of DRD4.
Most individuals carry 4 and/or 7 repeats of this 48 bp region, which can be easily genotyped
using PCR and gel electrophoresis. With this PCR, 4-repeats will generate a fragment of 475 bp
and 7-repeats will generate a fragment of 619 bp.

Below is a diagram of a 2% agarose gel that has already been run at 100 volts for 45 min. Ladder
(i.e., molecular mass ruler) was loaded in lane A and PCR product from three different
individuals was loaded in lanes B – D.
1. The ladder contains DNA fragments of 1000, 700, 500, 200, and 100 bp. On the diagram
below, label the 1000 and 100 bp fragments. (2 pts)
2. An individual homozygous for 4-repeats was run in lane B. Draw the PCR fragment(s)
for this individual on the diagram. (1 pt)
3. An individual homozygous for 7-repeats was run in lane C. Draw the PCR fragment(s)
for this individual on the diagram. (1 pt)
4. An individual heterozygous for 4-repeats and a less common 5-repeat copy was run in
lane D. Draw the PCR fragment(s) for this individual on the diagram. (2 pts)
1) In gel electrophoresis, smaller DNA
fragments will travel faster than larger
fragments since it is easier for smaller
fragments to make their way through the
pours inside the gel. Therefore, the 100 bp
ladder band will travel farthest from the wells
and the 1000 bp band will travel the least.
2) 4-repeats will give a band at 475 bp, which
is located slightly below the 500 bp ladder
band. Because this individual is homozygous
for the 4-repeat allele, both copies of his/her
autosome (i.e., one from the mom and one
from the dad) will give a 475 bp band and no
other bands will be visible on the gel.
3) Similarly, a homozygous individual for 7-
repeats will give a single band at 619 bp,
which is midway between the 500 and 700 bp
ladder bands.
4) A heterozygous individual for the 4-repeat
and the 5-repeat alleles will have one band at
475 bp and another at 523 bp (i.e., 475 + 48
= 523 bp for the 5-repeat allele).
1
5. List one of the two reasons why it is important to run a ladder with your PCR fragments on an
agarose gel. (2 pts)

The ladder acts as a size standard for comparison to your PCR fragments to make sure
you are amplifying the correct fragment (if the size of the fragment is known before
hand) or to estimate the size of your PCR fragment (if the size is not known before hand).

The ladder also allows you to determine if the gel ran properly in the case where no PCR
fragment bands are visible.

6. A student assembled his gel rig as diagramed below. Is this the correct orientation? What
would happen if he were to run his PCR product on the agarose gel assembled in this
orientation? (2 pts)

No, this is not the correct orientation.

Because DNA is negatively charged, the
PCR product would run toward the positive
electrode and off of the gel if this orientation
were used.

Make sure you answer all parts of the

question in order to get full credit.