10/5/18

Rational:

To do a plaque assay to check for the presence of arthrobacter phage. Also to compare the soil D metadata with the last two soil samples.

Procedure:

• Cleaned lab desk with CiDecon and ethanol
• Weighed enriched lysate- 15.7 g
• Put in the centrifuge for 5 min at 3,000 G
• Weighed weigh boat with dry soil- 3.5 g
  • Dry soil- 1.04 g
  • Percent water- 15.4%
• Percent soil types
  • 4 mL sand- 44.4%
  • 3 mL silt- 33.3%
  • 2 mL clay- 22.2%
• Put 2 mL LB broth in control and soil D TA
• Filtered enriched lysate to get FS lysate
• Put 10 ML FS lysate into .5 mL arthro
• Put 22.5 ML CACl2 1M into the control and soil D TA
• Added arthro and FS lysate to soil D TA
• Added 2.5 mL TA to control and soil D TA
• Poured on plates and waited 10 min
• Put plates in incubator at 26 C at 3:30
• Cleaned lab desk with CiDecon and ethanol

Conclusion:

The percent water is similar to soil B, but significantly lower than soil C. The soil type for soil D is loam which is similar to soil B (clay loam), but a little different than soil C (clay). Next lab I will chack my plaque assay for plaque. If there is no plaque then I will collect a new soil sample. If there is contamination then I will redo the plaque assay. If there is plaque than I will start purification.

Fig.7.D – This is the control plate for soil D plaque assay. This yellow circles indicate the location on bubbles on the plate.

Fig.8.D – This image shows the plate for soil D’s plaque assay and the yellow circles show the location of bubbles on the plate.

Fig.9.D – This image shows the falcon tube which indicates the amount (and percentage) of sand, silt, and clay in a 10 mL sample of soil D.