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Revision as of 10:00, 3 May 2013
| Experimental Chemistry
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To conduct gel electrophoresis to test the activity of the DNase I attached to the enzyme.
Electrophoresis Gel Preparation</u.
- Add 50 µL HPLC water to each of the lyophelized buffer/DNA samles. Try to get as much dissolved as possible.
- Prepare a 1% agarose gel with 0.25g Agarose and 25mL TAE Buffer (microwaved together for 40 seconds).
- Set in the electrophoresis tray with the comb for 20+ minutes.
<u>Running The Gel
- A DNA control for the gel was prepared by placing 1 uL of DNA in 4 uL of water with 1 uL of loading dye
- 5 uL of each sample was placed in a mini-pcr tube. 1 uL of loading dye was added to each tube
- 6 uL of previously prepared ladder + dye were placed in the first lane. The other lanes were then filled with the samples prepared.
- The lanes were loaded in the following order: 1) Ladder, 2) DNA alone 3) DNase I, 4) DNase I + 1 uL NHS Azide, 5) DNase I + 10 uL NHS Azide, 6) DNase I + 100 uL NHS Azide, 7) DNase I + 200 uL NHS Azide, 8) DNase I + 500 uL NHS Azide, 9) Hydrogel without DNase I, 10) DNA alone (repeated)
- Once the gel was loaded,
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