TEM Results:

Unfortunately, the pictures were not of high quality since the copper mesh TEM disc was exposed to uranyl acetate for 15 seconds longer than the protocol instructed, and the disc was bent causing it to begin to deteriorate. The following pictures were taken on the TEM.

Rationale:

Finish DNA extraction protocols so the extracted DNA can be used on the Nanodrop to find the concentration of DNA.

Procedure:

1. Re-suspended phage pellet with 0.5 mL of distilled water.
2. Added 2 mL of 37ºC DNA Clean Up Resin to phage pellet which then was equally distributed into two microcentrifuge tubes.
3. Centrifuged microcentrifuge tubes at 12,500xg for 3 minutes and removed supernatant.
4. Added 1 mL of 80% isopropanol to each microcentrifuge tube and re-suspended pellet.
5. Repeated steps 3 and 4 twice.
6. Added 1 mL of 80% isopropanol to each microcentrifuge tube and re-suspended pellet.
7. Filtered each microcentrifuge tube through one-column syringe.
8. Centrifuged columns at 12,000xg for 5 minutes.
9. Added 50 µL of 80ºC Elution Buffer to each column.
10. Allowed columns to sit for 1 minute before centrifuged at 10,000xg for 1 minute.
11. Performed a blank on the Nanodrop by placing 2.0 µL of Elution Buffer on the sensor, then used 2.0 µL of exacted DNA.

Observations:

• The second phage of which the picture was taken had a head length of 55 nanometers and a tail length of 128 nanometers.
• The pellet that formed after adding the resin and centrifuging was white and cloudy.
• The purified phage genomic DNA was place into a microcentrifuge tube labeled “KEA Ferranti 11/26/18 888.1.”
• Ferranti contained 888.1 ng/µL of nucleic acid.