Rationale:

To finish extracting the DNA from the pellet frozen on 11.26.18 and to determine the concentration of the DNA via nanodrop.

Procedures:

1. Defrosted the pellet and added .5mL of sterile water to redissolve the DNA into solution.
2. Added 2mL of 37˙C resin to the vial and mixed.
3. Split the contents of the vial into two microcentrifuge tubes.
4. Centrifuged at 12500g for 3 minutes, removed the supernatant.
5. Added 1mL of 80% isopropanol alcohol and shook to dissolve the pellet.
6. Repeated steps 4-5 two more times.
7. Added 1mL of 80% isopropanol alcohol and shook to dissolve the pellet.
8. Added the contents of both tubes to separate vacuum columns and vacuumed.
9. Dried each column filter by centrifuging for 5 minutes at 12,000g in another microcentrifuge tube.
10. Added 100µL of elution buffer to each filter and transferred to another microcentrifuge tube and centrifuged at 12000g for 1 minute.
11. Combined the contents of both tubes into one new microcentrifuge tube and labeled “NMN 11.28.18 Pippa”.
12. Used 2µL of the DNA to conduct a Nanodrop test

Observations/Data:

When redissolving my pellet several chunks remained undissolved. The nanodrop results were 138.92 for nucleic Acid(ng/uL) with an A280/A260 ratio of 1.922.

Conclusions/Next-Steps:

From the nanodrop test, it can be concluded that I have a fairly high concentration of DNA from the phage “Pippa”, which can now be used to run a gel electrophoresis test this week and to sequence the genome in the future.