11/12/2018

Objective

To pick and purify the phage samples and isolate a particular type of phage.

Procedure

1. 37 plaques were picked from 4th purification run and deposited in a microcentrifuge with 100μl of phage buffer.
2. After phages are mixed in the microcentrifuge tube, 10 μl of the extracted phage solution is transferred to a microcentrifuge tube with 90μl of phage buffer to acquire a 10^-1 serial dilution.
3. 10 μl of phage extract from 10^-1 dilution is transferred to a microcentrifuge tube with 90 μl of phage buffer
4. the top agar mixture was prepared for 4 plates.
5. 8 ml of LB broth was added to a conical vial
6. 90 μl of CaCl2 was added to the conical vial.
7. Three 0.5 ml of arthrobacter samples were enriched with the extracted phages, the 10^-1 and 10^-2 dilution for 15 minutes.
8. 10 ml of 2X TA was added to the conical vial.
9. 4.5 ml of the TA mixtures was added to each enriched sample.
10. the samples and the mixture were then plated on three agar plates.
11. 4.5 ml of TA mixture was plated on another agar plate for a control plate.
12. after 15 minutes, all the plates were placed inverted in the incubator.

Analysis

Due to low titer of the sample, more purification is required to isolate a particular type of phage. more lysate will be used in the future to have a plate with more plaques.