Case Of The Crown Jewels Template Page 26
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28Therefore, 328 µL of the purchased lambda DNA would be mixed with 672 µL of dH
O. This
2
mixture is the stock lambda DNA to be used later in the experiment. Store the lambda DNA at –15
to -25°C until use.
2
Lambda DNA cut with EcoRI
2.1
Determine the concentration of the Lambda DNA/EcoRI (see the label from the
supplier).
2.2
Using the formulas in section 1, dilute the Lambda DNA/EcoRI to a final concentration
of 0.175 µg/µL.
2.3
Store the diluted Lambda DNA/EcoRI at –15 to -25°C until ready to use.
3
Aliquoting student samples
3.1
Set-up seven microcentrifuge tubes for each pair of students doing the lab. Label and
add the following to each set of tubes:
15 µL of lambda DNA
Crime Scene DNA
15 µL of lambda DNA
Suspect 1
15 µL of lambda DNA/EcoRI
Suspect 2
15 µL of 10X Buffer (supplied with the enzyme)
10X Restriction Buffer
12 µL of HindIII
Enzyme
20 µL of loading dye
Dye
dH
O
1 mL of distilled water
2
3.2
These tubes are to be shared between two students. The tubes should be stored at –15 to
-25°C until ready to use.
4
Laboratory station set-up
4.1
Prepare the laboratory stations; two students can work at each station. Place the
following at each station:
Crime Scene Kit (with reagents)
Pipet tips for 2 – 20 µL pipet
Tube rack with empty tubes labeled Crime, Susp-1, and Susp-2
2 – 20 µL pipet
C a s e o f t h e C r o w n J e w e l s
P a g e 24
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