Research Question:

To find out how the presence of bacteriophages in the soil around red or white oak trees has a correlation with the health condition of oak trees.

Rationale:

A PCR run can identify if there are phage DNA present in the enrichment lysate, an accurate way to detect phages through their specific conserved gene that decides the length of their tails.

PCR on Sample (10.24.18):

Materials:

1. Sample DNA from enrichment lysate sample (10.24.18) and groupmate sample.
2. 2X PCR mix
3. ddH2O
4. Primers (AK, AL, AM, AN, AO, AP, AQ, AR, AS, AT, AU, AV)
5. Thermocycler
6. Microcentrifuge tubes
7. Centrifuge
8. Control DNA

Procedure:

1. Spin down 1 ml of Enrichment Sample Lysate (10.24.18) at 3000 g for 10 min.
2. Cook Sample Lysate for 15 min.
3. Added 4.5 ul of ddH2O to the 12.5 ul PCR mix repeat for three tubes.
4. Added 4ul of primers 1, 2, and 3 to three different tubes.
5. Added 2ul of both samples, at a total of 4ul.
6. Place the mix in the thermocycler for PCR.

Observations, Interpretations & Conclusions:

This time the PCR mix wasn’t preadded with DNA loading dye, and the Lysate sample size has been doubled, going from 1 ul per sample to 2 ul per sample, hoping the larger quantity of sample lysate would yield more DNA replicas.

Next Step:

The next step would be running a DNA gel electrophoresis to determine if there is DNA present in the sample lysates.