SEA Phage Lab Day 29
Day 29: 11/28/16 DNA Restriction Digest
Materials: 4 different enzymes, buffers, ddH2O, DNA, micro-centrifuge tubes, pipettes
Purpose: To cut my phage’s genome into a lot of pieces based on its DNA’s sequence.
Procedure:
- We first calculated how much DNA, enzyme, water, and buffer we needed to put in each tube.
2. I labelled 5 micro-centrifuge tubes with my table and the numbers 1-5.
3. First, I added the correct amounts of ddH2O in each of the micro-centrifuge tubes.
4. Then, I retrieved my DNA and put it in the incubator for 10 minutes at 37 degrees celsius.
5. While I was waiting for 10 minutes, I added the 10x buffer in tubes 1, 4, and 5.
6. After 10 minutes, I centrifuged the DNA for about 30 seconds.
7. Then, I added 5 micro-liters of the DNA is each of the five tubes.
8. Then, I put the DNA tube back in the ice.
9. Afterwards, I added the BSTE1 enzyme in tube 2, the SCaI2 enzyme in tube 3, the ECORI enzyme in tube 4, and the Hind III enzyme in tube 5.
10. I stored my micro-centrifuge tubes in the incubator at 3:52 pm.
11. Dr. Adair took out my micro-centrifuge tubes from the incubator at 6 pm, and she put them in the bottom drawer of the fridge.
Next Step: Run the gel.