UW Stout/Ethanol FA19

Yeast torture 1

Protocol:

Materials:

1. neat ethanol
2. yeast
3. sterile water
4. 200 ul
5. 96 well plate

Procedure:

1. (add 50ul mixture to 50ul yeast in 96 well plate.)x2. Used 12 wells in total
2. fill 6 200ul tubes, each with water and ethanol concentration x2. Resulting in 6 200ul tubes with 100ul of mixture in each.

	1	2	3	4	5	6
ul ethanol	0	3.75	7	10.25	13.5	16.75
ul water	50	46.25	43	39.75	37.5	33.25
ul yeast	50	50	50	50	50	50

Knockout Strand Yeast Torture

Protocol:

Materials:

• Yeast Knockout Strains:

1. 1. YAL061W
   2. YBL086C
   3. YBL9081W
   4. YBR033W
   5. YBR138C
   6. YBR220C
   7. YHR048W
   8. YGL140C
   9. YJR061W
   10. YKR005C
   11. WT:BY4735

• neat ethanol
• sterile water
• micropipeter(s)
• 96 well plate

Procedure:

1. add 50 ul of Knockout Strain to individual well (96 well plate).
2. add 50 ul of water+ethanol mixture to individual well (Follow table below).

Well	1	2	3	4	5	6	7	8	9	10	11	12
ul yeast	50	50	50	50	50	50	50	50	50	50	50	50
ul ethanol	7	7	7	7	7	7	7	7	7	7	7	7
ul water	43	43	43	43	43	43	43	43	43	43	43	43