November 30

11/28 ~ DNA extraction

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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:

The spot test to confirm the strength of the titer (Was 1 X 10^9)

 

Conclusion/Next Steps: Will be continuing the next steps of DNA extraction with the hopes of breaking open the capsid of the bacteriophage.


Posted November 30, 2018 by justin_yu1 in category Justin Yu

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