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 spot test for the diluted samples would be a fast way to determine the best sample volume to web and flood the plate and get the most phages possible.

Experimental Procedure for Spotting Test for Soil (Gabe):

(1) Lysate

(2) LB Broth

(3) 2x TA

(4) 1M CaCl2

(5) Arthrobacter host

(8) Phage Buffer

1. The plate was flooded and then lysate filtered.

2. The lysate was diluted into 10^0, 10^-1, 10^-2, 10^-3, 10^-4, 10^-5, 10^-6, 10^-7, 10^-8.

3. The top agar was prepared with 2.0 ml LB broth, 0.5 ml Arthrobacter, 4.5 mM CaCl2 and 2.5 ml 2X TA.

4. The top agar was poured onto the plate and set still to solidify for 15 min.

5. The lysate dilutions were then spotted onto their respective sections on the plate and set still 15 min

6. The plate was then placed in incubator for culture.

Observations, Results & Data:

The plate was almost completely lysed, only traces of colonies can be found on the sample plate.

Interpretations & Conclusions:

Due to the plate being almost completely lysed, the plate could then be flooded and collect the lysate.

Next Step:

The results of the spot test would help determine the dilution optimal for webbing a plate.