November
30
11/28 ~ DNA extraction
Rationale: Will be running the procedure of pelleting the lysate to obtain the bacteriophage DNA
Procedure:
- Created an aseptic zone to prevent bacterial contamination
- Added 10mL of lysate into a 50mL conical tube
- Pipetted 40μL of nuclease into the tube and inverted the tube around 10 times
- Added in 4mL of PEG and inverted the tube one to two times
- Placed the tube into the shaker for 30 minutes
- Took the tube out of the shaker and allowed to sit in room temperature for 45 minutes
- After the allotted time, moved the tube into a centrifuge and allowed to centrifuge for 20 minutes at 10,000 G
- After centrifuging, decanted the supernatant in the tube and collected the pellet(s)
Observation:
Conclusion/Next Steps: Will be continuing the next steps of DNA extraction with the hopes of breaking open the capsid of the bacteriophage.