Rationale: Flooded a mostly lysed plate and creating a webbed plate from previous lab’s calculations

Procedure:

• Created an aseptic zone to prevent contamination of plates by other bacteria strains
• Obtained the lysed plate from refrigeration and obtain a 50mL conical vial of PB
• Pipetted 6mL PB into the plate and allowed to swirl on a shaker for 1 hour and 15 minutes
• During the waiting period, obtained a 50mL conical vial and pipetted 6mL of LB Broth and 76.5mL CaCl2
• Added 5.25μL of lysate into 0.5mL arthrobacter and 10μL of lysate into another 0.5mL of arthrobacter
• Pipetted 7.5mL 2xTA into the 50mL vial and pipetted 4.5mL into each arthobacter + lysate tube (Respectively)
• Immediately plated and allowed to sit for 15 minutes
• After the allotted time, moved the plates into incubation
• After the hour and 15 minutes passed, removed the flooded plate from the shaker and used a syringe filter and a 22μL filter to filter the lysate
• Placed the new lysate (Lysate 5) into refrigeration

Observations:

The 10μL webbed plate after pouring

The 5.25μL plate after pouring

Next Steps/Conclusion: During the next lab, if the webbed plates turned out lysed, will be flooding the two plates and combining their lysates to create lysate 6. After getting the lysate, will be calculating their strength by running serial dilutions