Discovery of Gibbonz
September 4, 2017
Collecting Environmental Samples
- I stopped at three locations on the highway between Stephenville and Granbury and collected soil samples
- Followed Protocol 5.1: Collecting Environmental Samples
Sample | 1 | 2 | 3 |
Date of Collection | 9/4/2017 | 9/4/2017 | 9/4/2017 |
Sample Type | soil | soil | soil |
Location Description | Fence line | Pond over flow | Road drainage |
General Location | Paluxy River | Stock Tank | Drainage |
Specific Location | 92665 80088 | 97471 83192 | 03093 84599 |
Sample Depth | 1.5 inches | 1 inch | 0.5 inches |
Ambient Temperature | 88°F | 88°F | 89°F |
September 6, 2017
Isolation of PHAGES from samples
- Followed Protocol 5.2: Direct Isolation, then completed Protocol 5.3: Plaque Assay
- I left the plate in the incubator for 48 hours. No phages were present.
- Followed Protocol 5.5: Enriched Isolation – Day 1
- I let the enriched culture incubate for 5 days
September 11, 2017
Continued Isolation of PHAGES
- Continued Protocol 5.5: Enriched Isolation – Day 2
- Followed Protocol 5.6: Spot Test
- I let the filtrate sit on the top agar for 15 min
- I inverted the plate for a second prior to incubation (caused filtrate to move a little)
- Incubated for 48 hrs
September 13, 2017
Checked Spot Test #1 and Started Spot Test #2
- No phage present for spot test number one of Enriched sample (See images)
- Followed Protocol 5.6: Spot Test
- Tried drying condensation on top of agar plate with Kimwipe
- Borrowed Enriched sample from Travis #3 in quadrant 2
- Borrowed Enriched sample from Mario #2 in quadrant 6
- Let filtrate sit for 20 min before incubating for 48 hrs
September 15, 2017
Checked Spot Test #2
- Spot test #2 was so contaminated that Dr. Edwards removed sample from incubator
September 18, 2017
Adopting a PHAGE
- Adopted a plaque from Morgan’s Direct Sample (See images “JM”) using Protocol 5.4: Picking a Plaque
Sample 3 Date of Collection 9/4/2017 Sample Type soil Location Description compost bed General Location Subdivison Specific Location 29° 34′ 36.480″ N, 98° 15′ 54.360″ W Sample Depth 1.5 inches Ambient Temperature 82°F - Then followed Protocol 6.2: Serial Dilutions
- I did dilutions 10^-1 through 10^-6
- On 10^-1 I used 90 micro-liters instead of 10 micro-liters
- Then followed Protocol 5.3: Plaque Assay for each dilution.
- Incubated for 48 hrs
September 20, 2017
Results of First Dilution and Start Second Dilution
- Results (See images)
- 10^-1: Lots of plaques, uncountable
- 10^-2: Approximately 35 plaques
- 10^-3: 5 plaques present
- 10^-4—-10^-6: no plaques present
- Chose isolated plaque from 10^-3 plate using Protocol 5.4: Picking a Plaque (See images)
- Then followed Protocol 6.2: Serial Dilutions again
- I used 10 micro-liters on all dilutions this time
- I did dilutions 10^-1 through 10^-6
- Then followed Protocol 5.3: Plaque Assay for each dilution
- Incubated for 48 hrs
September 22, 2017
Results of Second Dilution
- Results (See images)
- 10^-1: Lots of plaques, uncountable
- 10^-2: Over 100 plaques
- 10^3: 3 plaques present
- 10^-4—-10^-6: no plaques present