November
30
DNA Extraction and Nanodrop Lysate 8
11/28/18
Rational:
To do DNA extraction on lysate 8 in order to obtain a sample of DNA from the phage found in lysate 8.
Procedure:
- Cleaned lab desk
- Added 500 ML of sterile water to phage precipitate
- Added 2 mL of DNA clean up resin and mixed
- Put the mixture into two centrifuge tubes and spun at 12,500 G for 3 min
- Pulled off the supernatent
- Added 1 mL of 80% isopropenol and spun at 12,500 G for 3 min
- Pulled off the supernatent
- Repeated the previous two steps two more times
- Added 1 mL of 80% isopropenol
- Put the solution in a column tube and filtered
- Centrifuged the filtered column at 12,000 G for 5 min
- Put the remaining solution in a new centrifuge tube and added 100 ML of 80 C elution buffer
- Centrifuged for 1 min at 12,000 G
- Ran the nanodrop on the sample
Observations:
- DNA concentration- 417.74 ng/ML
Conclution:
The nanodrop results showed the the DNA concentration in lysate 8 was 417.74 ng/ML. As there is not enough time nothing else will be done with this sample however, next I would have done PCR and gel electrophoresis on lysate 8 since it has a high titer.