UW Stout/Lithium chloride FA19

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Safety Precautions:

~Safety Goggles

~Gloves

~Lab Coat

~Work under a chemical hood

Materials:

~96 plate

~Yeast strands

~H20

~Lithium Chloride

~250ml beaker

~10, 1.7ml microcentrifuge tube

Protocol:

Create a sufficient amount of 10μM stock solution, mix 72μl of sterile H2O with 8μl of 100μM stock solution, in order to replicate series 9 from the previous experiment. Add 45μl of sterile H2O to each of the wells. Add 5μl of diluted caffeine solution to each well. Then add 50μl of each strand of yeast being tested. After the solution in the wells is completed put the 96 well plate into the SpectaMax Plus 384 Microplate Reader where the data will be collected every 5 minutes for the next 24 hours.


Protocol:

1. Make a solution of lithium chloride with 4.24g of LiCl and 9mL of H20, then adjust the volume to 1mL of H20 in a 100ml beaker

2. Filter sterilize with .45 micron syringe filter

3. Make 11 different concentration in 1.7ml microcentrifuge tubes with 20ul of H2O starting at 0.5% concentration increasing to a 5.5% concentration

4.Pipette 5 u from solution 1 into the second well on the 96 plates, and the second solution into the second well, until each solution is in each well.

5. In the first well add 10ul of H20 without any LiCl solution

liclconcentraion!.png


Materials

~Sterile H2O

~Yeast Strands

~96 Well Plate

~LiCl @ 1M concentration