October 19

PCR 10/17/2018

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Rationale: To conserve resources, we ran a pcr test on our enriched isolations to check for the presence of phage DNA.

Process:

  1. Wash table
  2. centrifuged enriched isolation
    1. 5 minutes at 3000 G
  3. added ~ 1 mL to microcentrifuge tube
  4. gave tube to Lathan to boil
  5. Prepared 4 different PCR reactions as shown in the table below
    1. Reagents Tube 1 Tube 2 Tube 3 Pos. Control
      TAQ Polymerase 12.5 µL 12.5 µL 12.5 µL 12.5 µL
      Primer Mix 4 µL PM1 4 µL PM2 4 µL PM3 4 µL PM1
      DNA RSM 1 µL 1 µL 1 µL NA
      DNA SJ 1 µL 1 µL 1 µL NA
      Pos. Control DNA NA NA NA 1 µL
      dd H2O 6.5 µL 6.5 µL 6.5 µL 7.5 µL
  6. Gave tubes to Lathan to run PCR cycle in PCR machine

Next steps: Run PCR products on a gel electrophoresis to check for bands that could indicate phage DNA. Also, calculate % sand/silt/clay.


Posted October 19, 2018 by rachel_melone1 in category Rachel Melone

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