11/16/18

Rational:

To estimate the titer of lysate 8 in order to determine whether a new lysate need to be obtained or not.

Procedure:

• Estimated titer from the spot titer using the 10^-7 dilution
• Titer= (5 pfu/10 ML)(1000 ML/mL)(10^7 ML/mL)

Observations:

• The control plate showed signs of contamination though the spot titer did not
• Most of the spots were cleared
• The spots for 10^-7 and 10^-8 were the only ones that were not cleared
• The 10^-8 spot showed no signs of plaque
• The arthro also seemed to be growing back over the cleared spots
• Titer= 5*10^9 (high titer)

Conclusion:

Since the spot titer showed a high titer then I will do a plaque assay on the 10^-7 dilution in order to calculate the actual titer. I will also do a TEM with this lysate since it is a high titer.

Fig.1.8 – This figure shows the results of the spot titer. The image shows that all of the dilutions up to 10^-7 were cleared. It also shows that there were no plaque for the 10^-8 dilution.

Fig.2.8 – This image shows the contamination found on the control plate for this test.