November 2

Spot Test 10/31/2018

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Rationale: Complete spot test with new sample isolations and the re-purified products from the heating and killing bacteria method to check for plaques.

Process:

  1. Set up aseptic system
  2. Made LB agar media for 1 control plate and 2 sample plates
    1. Reagents Control Sample
      LB Broth 2 mL 4 mL
      2x TA 2.5 mL 5 mL
      CaCl2 23 µL 45 µL
      arthro NA 1 mL
  3. poured 5 mL of LB agar media into each plate and let sit ~5 min
  4. filtered enriched sample with syringe filter
  5. pipetted 5 µL of direct, enriched, RSM1, SJ1, LCG1, RSM2, SJ2, LCG2, and phage buffer (negative control) into designated areas of sample plate and let sit ~10 min
    1. the initialed samples are the results from the heating and killing bacteria method
    2. the 1 indicates arthro was added and the 2 indicates arthro was not added (except in the case of LCG2, where arthro was added)
  6. incubated for 48 hours

Next steps: If we get plaque we may try to get a high titer lysate  from our own sample, but we will most likely just use a classmate’s lsyate and start to work towards getting higher titer by first performing a plaque assay.


Posted November 2, 2018 by rachel_melone1 in category Rachel Melone

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