11/7/18

Rational:

To get a lysate that will give a high titer using 5 plates instead of one since the 5 plates prepared last lab are not usable.

Procedure:

• Cleaned lab desk with CiDecon and ethanol
• Put 180 ML of PB to two microcentrifuge tubes
• Added 20 ML of lysate to both
• Added 20 ML of 10^-2 lysate to 5 tubes of .5 mL arthro
• Waited 10 min
• Put 12 mL of LB broth in a tube
• Added 135 ML CaCl2
• Added 2 mL of the TA mixture to the 5 tubes of arthro
• Added 2.5 mL to the 5 tubes and the control and poured them on the plates
• Let the plates sit for 10 min
• Put in the incubator at 26 C at 3:20

Observations:

• Two of the plates slipped though one was still webbed
• One of the plates was completely cleared
• The other two plates were webbed

Conclusion:

Since three of the five plates were unusable the 5 more plates were prepared. Next lab we will flood the 5 plates in order to get a new higher titer lysate. A plaque assay will then be done in order to calculate the new titer.

Fig.2.CW – This image shows one of the plates that slipped, however the majority of the plate still shows the results that were expected.

Fig.3.CW – This image shows one of the plates that was webbed, but did not slip of completely clear the plate.

Fig.4.CW – This image shows the other plate that slipped, though there is no signs of any arthro growing on it.