To get deletion collection mutants from the -70 freezer:

(August 2006)

1. Go to www.yeastgenome.org and get the ORF name of the gene deletions you want.

2. Go to the Depressed, shared, Young_Lab_iMac_shared, databases", and open "2004mat_a_obs_v1.0.xls" or "2004mat_alpha_obs_v1.0.xls". Go to the worksheet tab at the bottom of the screen called "mat_a_obs" or "mat_alpha_obs". Find your gene and write down the information in columns E, F and G (plate, row and column). E has the number of the 96-well microtiter plate. F and G have the row and column within the microtiter plate.

3. Assemble the following:

• dry ice in a container large enough to hold a 96-well microtiter plate

• one sterile 18 gauge needle (pink) for each strain you are collecting

• one square piece of thermowell aluminum sealing tape (costar 6569), ~1 x 1 cm, for each strain you are collecting (as of August 2006, sealing tape is in the drawer 94, Chris's bench)

• YPD plates

• sterile toothpicks

• ethanol and kimwipes

4. Wipe the work area with ethanol. Wear gloves and keep everything as sterile as possible.

5. Delection collection boxes are on the top shelf of the upright -70 in J434. Use the ones in the white, hand-labeled freezer boxes. Take out the microtiter plate you need and put it on dry ice.

6. Take off the plastic lid and puncture through the foil over the well with your strain, using the sterile 18 gauge needle. Dig into the ice and take as large a chunk as possible to transfer to the YPD. Take out more with a toothpick until you have a large droplet on the plate.

7. Cover the well again with the square of aluminum sealing tape. Replace the plastic lid and put the plate back in the freezer.

8. Streak out the drop of cells on the YPD.