BIOL150 Montgomery College, Rockville Sizing DNA on an agarose gel
This assignment is due by 07/03/2017. Late submissions will be penalized (25%
deduction/day). This assignment is worth 15 points. This assignment may not be
submitted electronically.
Agarose gel electrophoresis can also be used to determine the size of a particular DNA
fragment. Whenever this technique is performed, a molecular size marker (also called DNA
Ladder) is used as a standard. The ladder contains DNA fragments of known size. Since
these fragments travel down the gel according to their size, a standard curve of the
distance traveled vs. fragment size can be drawn for the DNA fragments of the ladder.
Scientists can then use that standard curve to estimate the size of unknown DNA
fragments.
In this assignment you will:
1. Create a standard curve of DNA ladder fragments and their migration distance by
graphing the given data.
2. Estimate the size of unknown DNA fragments using the standard curve
Build the standard curve: Using the gel represented in Figure 1 measure the distance traveled (in millimeters)
by each fragment in the DNA ladder (Lane 1). Start measuring from the center of
the well (open rectangle) and measure the distance to the center of each band.
Enter your data in Table 1. Plot the length of each fragment on the Y axis versus distance traveled on the X
axis. Use the graph provided. fragment length is measured in base pairs or bps distance traveled is measured in millimeters or mm Y axis is the vertical axis X axis is the horizontal axis Draw a best fit curve. To do this, draw a smooth curve * that best estimates the
plotted data. Do not just connect the dots. It is ok if your curve misses some of the
data to maintain a smooth curve. Estimate the size of unknown DNA fragments: The unknown DNA fragments are in Lanes 2 to 4. Measure the distance traveled by
each unknown the same way you did for the Ladder. Record your data in Table 2. Use your standard curve to estimate the size of the unknown fragments (In lanes 2,
3 and 4).
o Mark the distance traveled by each unknown fragment on the X axis. From
each point, draw a line up until your line touches the standard curve, then
make a horizontal line over to the Y axis. This will give you the size of the
fragment of interest. (show your work on the curve) BIOL150 Montgomery College, Rockville Please submit only your report page (your standard curve as well as Tables 1 and 2).
Your report must be neat. Points will be deducted for sloppiness. BIOL150 Montgomery College, Rockville Figure 1: Agarose gel with DNA Ladder in Lane 1, and unknown DNA fragments in
Lane 2-4. Empty boxes are the wells, the solid bands are the DNA fragments
after electrophoresis. Take measurements in millimeters (mm). ** Example of a smooth curve BIOL150 Montgomery College, Rockville Report Page Name: Table 1. Distance traveled by the fragments of the DNA Ladder (Lane 1).
Molecular weight
marker
fragment size (bp)
2,000
1,500
1,000
750
500
300
150
50 Distance traveled
by the fragments
(mm) Table 2. Distance traveled by the fragments in lanes 2 to 4. Lane
2
3
4
Standard Curve: Distance traveled
by the fragments
(mm) Size of the
fragments
(bp) BIOL150 Montgomery College, Rockville
2500 2000 1500
length of DNA fragment (base pairs)
1000 500 0 0 10 20 30 40 50 60 70 80
distance (mm)

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