Week 02 - Yeast mating

Submitted by kdorfman on Thu, 07/20/2023 - 18:16

Yeast labs, week of 9/11/23 (2nd week of classes, 1st complete week)

See notes here

Total YED plates for the week for 4 sections plus prep: 75

Students mate 3 A mating types with 3 alpha mating types

Take YED plates out early so they stop sweating.

Make an assembly line of 7 stations (use templates):

station activity materials
1 label plate YED plates, labeling template, Fisher Finest marker
2 plate HA1 across 3 small plates of HA1 on YEPAD, sterile toothpicks, used toothpick bucket
3 plate HA2 across 3 small plates of HA2 on YEPAD, sterile toothpicks, used toothpick bucket
4 plate HAR across 3 small plates of HAR on YEPAD, sterile toothpicks, used toothpick bucket
5 Plate HB1 down, mixing carefully with the A strain already there, new toothpick each time 3 small plates of HB1 on YEPAD, sterile toothpicks, used toothpick bucket
6 Plate HB2 down 3 small plates of HB2 on YEPAD, sterile toothpicks, used toothpick bucket
7 Plate HBR down 3 small plates of HBR on YEPAD, sterile toothpicks, used toothpick bucket

For mating, students need these strains on YEPAD plates so they all start out white:

  • HA1
  • HB1
  • HA2
  • HB2
  • HAR
  • HBR

For observation, students need these strains on YED plates so their colors can be observed:

  • HA0
  • HB0
  • HA1
  • HB1
  • HA2
  • HB2
  • HAR
  • HBR

For observation of colors, make demo plates (maybe 4 of each):

  • 4 YED plate with 4 A strains
  • 4 YED plate with 4 alpha (B) strains

(If you let the parental strains sit on YED more than a couple of days, the mutant strains turn red, and the red color will still be there even after there are heterozygous diploids with complementation.)

List of yeast strains

A types alpha (B) types
HA0 (WT) HB0 (WT)
HA1 (ade1) HB1 (ade1)
HA2 (ade2) HB2 (ade2)

Start 6 strains at least the Friday before the first lab.

Instructors should make some mating plates to have in the fridge to hand out when student plates are bad.