Title: Metadata, Spot Test, and Plaque Assay

Date: September 12, 2018

Rationale: The sand-silt-clay results and the % water results are ready, and the lysate is done incubating, therefore metadata can be recorded, and tests be done to check for plaques.

Procedure: Aseptic zone created by the following procedure:

• Counter washed and wiped with CiDecon
• Counter sprayed with 70% EtOH and allowed to evaporate completely (to dehydrate and kill any bacteria on the counter and avoid contamination)
• Ethanol lamp lit to create rising heat and a current that protests samples from falling contamination.

% Water: The dirt weighed 5.37 grams before evaporation and weighed 5.066 after evaporation. The following calculation yields the % water of the collected soil:

1-(5.066/5.37)= .057 = 5.7% water

Sand – Silt – Clay: 3.25 mL of soil was layered:

1 mL of sand      1.75 mL of silt   .5 mL of clay

= 30.77% sand     =53.85% silt    = 15.38% clay

1. The enriched lysate from Monday was pulled from the incubator, massed at 21.47g, and spun in a centrifuge to separate cells.
2. A spot test and a plaque assay were done to assure the best chances of a phage discovery.

Spot Test                                                                              Plaque Assay (All multiplied by 4 for 3 plates + negative control)

– 2.0 mL LB Broth                                                                            – 2.0 mL LB Broth

– 0.5 mL Arthrobacter                                                                    – 0.5 mL Arthrobacter

– 2.5 mL 2X Top Agar                                                                      – 2.5 mL 2X Top Agar

– 22.5 microliters CaCl₂                                                                  – 22.5 microliters CaCl₂

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~ 10 microliters spotted onto plate –> put into incubator                       Set 15 min –> put into incubator