11/14/18

Rational:

To do a spot test titer from the lysate obtained from last lab’s serial dilution in order to calculate the new lysate’s titer.

Procedure:

• Cleaned lab desk
• Filtered the flooded plates from last lab
• Put 90 ML of PB into 8 microcentrifuge tubes
• Diluted the lysate to get 10^1, 10^-2, … and 10^-8 dilutions
• Put 2 mL of LB broth into a tube
• Added 22.5 ML CaCl2, 2.5 mL TA, and .5 mL arthro
• Poured on plate and waited 10 min
• Put 2 mL of LB broth into a tube for the control
• Added 22.5 ML CaCl2 and 2.5 mL TA
• Poured on plate and waited 10 min
• Spotted the dilutions and lysate onto the plate and waited 10 min
• Put in incubator at 26 C at 4

Conclusion:

The plates from lysate 7 had been completly cleared after 24 hrs so they had been flooded the day before in order to prepare a new lysate. Next lab the titer of this lysate will be determined using the spot titer.