Title: PCR & Metadata Results

Date: 22 October 2018

Rationale: A PCR (Polymerase Chain Reaction) will be done to attempt to replicate any phage DNA present in the most recently acquired soil sample. The metadata results will also be recorded and interpreted.

Procedure:

• The enriched lysate from the previous lab session was centrifuged at 3000 G for 5 minutes to separate the Arthrobacter from the phage and the lysate suspension.
• ~1 mL of the centrifuged lysate was placed into a microcentrifuge tube and boiled to release capsid DNA from any potential phage
• 3 different primer mixes with different DNA primers were made using the following recipe:
  • 12.5 microliters of Taq Polymerase
  • 4 microliters of primer mix (PM1, PM2, or PM3)
  • 6.5 microliters ddH2O
  • 2 microliters of DNA (1 microliter of boiled lysate + 1 microliter of a partner’s lysate)
• The mixture is put onto ice and the results will be checked with a gel in the following lab
• The % water and soil dipsersion results were checked and recorded

Results/Conclusions: The final mass of the soil sample was 9.437 g meaning that the % water composition of the soil was 16.19% H20. As for the soil dispersion, the soil was composed of 42.86% Sand, 28.57% silt, and 28.57% clay. The PCR mix is able to recognize phage DNA and, because of the primer mixes, bind to the DNA strands and replicated any phage DNA that are present. This provides another method to test phage presence as opposed to a plaque assay or a spot test.