9/26/18

Rational:

A new plaque assay will be performed as the last plaque assay was not done with arthrobacter and some other bacteria instead. So a new plaque assay will be performed in order to check for the presence of arthrobacter phage.

Procedure:

• Cleaned lab desk with CiDecon and ethanol
• Set up and aseptic zone
• Put 10 ML FS lysate into 400 ML arthro
• Put 2 mL LB broth into TA mixture
• Added 22.5 ML 1M CaCl2 to the TA mixture
• Added another .1  mL LB broth to the TA mixture
• Added 400 ML arthro and lysate to the TA mixture
• Added 2.5 mL TA
• Poured on plate and waited 10 minutes
• Put 2.1 mL LB broth to control TA mixture
• Added 22.5 ML 1M CaCl2 to control
• Added 2.5 mL TA to the control
• Poured on the plate and waited 10 minutes
• Put plates in incubator at 26 C at 3:30
• Cleaned lab desk with CiDecon and ethanol

Conclusion:

The plates from 9/24 were not able to comfirm the presence or absence of arthrobacter phage in soil C. This is because the bacteria that was used was not the arthrobacter that has been used, so any plaques seen would be from the presence of of a different type of phage. Contamination was also seen on both the control and plaque assay plates this in itself would have required that I new plaque assay be done. Next lab I will check for plaque on plaque assay soil C 2. If there is plaque then I will start purification and if there is not then I will collect a new soil sample and filter it and collect metadata. If there is contamination I will do another plaque assay.

Fig.7.C – This image shows the plaque assay for soil C                   Fig.8.C – This image shows the control plate for soil    (done with an unknown bacteria not arthrobacter). The               C which also has an unknown bacteria growing on it.  white spots show contamination.                                                        This plate is covered in white spots indicating                                                                                                                                  contamination (may be difficult to see).

Fig.9.C – This shows the control plate for plaque                            Fig.10.C – This image shows the plate for soil C        assay soil C 2. The yellow circles in indicate where                         plaque assay 2. The yellow circle shows where a          bubbles are located to prevent them from being                             bubble is to prevent it from being mistaken as a          mistaken for plaque.                                                                              plaque.