Rationale

New titer and web calculations were made since plate was not webbed at all. Made more webbed plates for flooding and serial dilutions on next lab day.

Detailed Procedures

1. Took 0.5 mL Arthro and added 203 microliters of 10^0. Sat for 15 mins.
2. Took another 0.5 mL Arthro and added 5 microliters of 10^0. Sat for 15 mins.
3. Took 6 mL of LB Broth, 7.5 mL 2X TA, and 69 microliters of 1 M CaCl2 into a 50 mL vial.
4. Took 4.5 mL of top agar mixture and poured onto a plate labeled as “control.”
5. Took another 4.5 mL top agar mixture and poured into a 15 mL vial labeled as “203 microliters.”
6. Took another 4.5 mL top agar mixture and poured into a different 15 mL vial labeled as “5 microliters.”
7. Took mixture from step 1 and added to 15 mL vial labeled as “203 microliters.”
8. Took mixture from step 2 and added to 15 mL vial labeled as “5 microliters.”
9. Took “203 microliter” vial and poured onto “203 microliters” plate.
10. Took “5 microliter” vial and poured onto “5 microliters” plate.
11. Let all three plate solidify for around 15 mins and inverted into incubator.

Results

Current titer= 1.4 x 10^7

web= 2.02 microliter of 10^0

Made a miscalculation and did 203 microliters of 10^0, so made another plate with 5 microliters of 10^0  instead of 2.02 microliters.

Conclusion/Next Steps

Plates from lab day 28 did not have any contamination. Since 10^-5 lysate was used, the plaque size was very small in size and quantity, that is why a new titer calculation was made from that. 2.02 microliters of 10^0 was small, so I was advised to used 5 microliters instead. Hopefully a webbed plate will be ready by next lab day so that flooding and serial dilutions will take place and get a high titer by Friday.

results from lab day 28