Case Of The Crown Jewels Template Page 25
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28Case of the Crown Jewels
Laboratory Preparation
NOTE: The following information contains instructions to order the reagents and supplies needed to prepare the
solutions and perform this laboratory activity.
Materials
Catalog
Item
Company
Phone Number
Quantity
Number
Lambda DNA
Fisher Scientific
800-955-1177
CDS1120216
80 µg
25 µg
Lambda DNA/cut with
Fisher Scientific
800-955-1177
S1120220
EcoRI
HindIII with 10X Buffer
Fisher Scientific
800-955-1177
BP3372-1
5000 units
Loading Dye
Fisher Scientific
800-955-1177
BP645-1
1 mL
Crime Sample Preparation
1
Lambda DNA
1.1
Place the tube of Lambda DNA purchased from the supplier in a 65°C heating block for
five minutes. Remove and invert several times to mix. Allow the tube to cool to room
temperature. Determine the concentration of the Lambda DNA (see the label on the
tube). The final concentration of the lambda DNA should be 0.175 µg/µL. Calculate
and perform a dilution using the following formula:
(Final concentration) X (Final volume) = (Initial concentration) X (Initial volume)
0.175 µg/µL
Final concentration
1000 µL
Final volume
Initial concentration
Concentration of the lambda DNA from the supplier
Initial volume
Total volume of the lambda DNA from the supplier
For example:
(0.175 µg/µL) (1000 µL)
X =
(Concentration of lambda DNA from the supplier)
Where X = volume of the lambda DNA that will be diluted to 0.175 µg/µl in a final volume of 1000
µL with distilled water (dH
O).
2
1.2
After determining the amount of lambda DNA from the supplier to use, subtract that
amount from 1000 to determine the total volume of water to add.
For example - If the tube of lambda DNA has a concentration of 0.533 µg/µL of DNA, then the
calculation would be as follows:
(0.175 µg/µL) (1000 µL) = (0.533 µg/µL) X
(0.175 µg/µL) (1000 µL)
(0.533 µg/µL
X =
X = 328 µL
Y = 1000 µL – 328 µL
Y = 672 µL dH
O
2
C a s e o f t h e C r o w n J e w e l s
P a g e 23
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