9/24/18

Rational:

To do a plaque assay in order to see if soil C has any arthrobacter phage. Record results results of soil C metadata in order to compare with soil B metadata.

Procedure:

• Cleaned lab desk with CiDecon and ethanol
• Dry soil C with weigh boat- 5.25 g
  • Dry soil C- 2.93 g
  • Percent Water- 47.9%
• Sand- 3.5 mL     Silt- 2.5 mL     Clay- 4 mL
  • Percent Sand- 35%     Percent Silt- 25%     Percent Clay- 40%
• Cleaned syringe filter to prevent contamination
• Filtered enriched lysate with the syringe filter
• Put 10 ML FS lysate into .5 arthro
• Put 2 mL LB broth into control and plaque assay TA mixtures
• Added 22.5 ML 1M CaCl2 to TA mixture for soil C plaque assay and control
• Added .5 mL arthro and FS lysate to plaque assay TA mixture
• Added 2.5 mL TA to TA mixture for soil C and control
• Poured onto soil C plaque assay and control plate and let sit dor 10 minutes
• Put plates in incubator at 26 C at 3:40-2:30 9/26

Conclusion:

The soil C metadata showed that soil C was clay according to the soil texture triangle. This is a slightly different soil type than soil B which was clay loam although the percent sand and silt change by only 5% and clay change by 10%. On the other hand the percent water in soil C (47.9%) is significantly greater than soil B (16.2%). Next lab I will check for plaque if there is a plaque then I will start purification. If there is contamination then I will redo my plaque assay and check for plaque again. If there is no plaque then I will get a new soil sample and filter it and collect metadata.

Fig.5.C – This shows the plaque assay for soil B. The                 Fig.6.C – This image shows the control plate for soil C.    yellow circles indicate the location of bubbles in the                 The yellow circles on the image indicate the location of  agar to prevent them from being mistaken as plaque.               bubbles that could later be mistaken as plaque.