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YPD FILLED MICROTITER PLATES

Uses:

1. Grow individual strains to use for PCR
2. Shipping strains

1. Make YPD regular agar (1.5-2%) for sending strains.

2. Make 200ml batches = enough for 6 microtiter plates.

3. Add G418 at 200mg/L.

4. Keep the medium at 65° or hotter. Place the multichannel pipettor basin on a block
that is set at 65°. Pour the hot medium nearly to the top.

5. Dispense agar media into wells. Do not blow out the final drop of
medium into the wells (makes bubbles).

Colony PCR: 200µl per well

Prior to inoculating cells, add 10µl water to each well. Then inoculate the
wells with sterile toothpicks. After all the wells are inoculated, give the
plate a gentle swirl to spread the cells on the surface of the medium in
each well. This makes semi-liquid cultures for use as template for
colony PCR, after which the plates can be parafilmed and stored.

Sending strains: 150µl per well

Let plates sit on bench to dry for several days before inoculating wells.
Do NOT add water to the wells.

from Linda Riles, November 1997



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		YPD FILLED MICROTITER PLATES Uses: 1. Grow individual strains to use for PCR 2. Shipping strains 1. Make YPD regular agar (1.5-2%) for sending strains. 2. Make 200ml batches = enough for 6 microtiter plates. 3. Add G418 at 200mg/L. 4. Keep the medium at 65° or hotter. Place the multichannel pipettor basin on a block that is set at 65°. Pour the hot medium nearly to the top. 5. Dispense agar media into wells. Do not blow out the final drop of medium into the wells (makes bubbles). Colony PCR: 200µl per well Prior to inoculating cells, add 10µl water to each well. Then inoculate the wells with sterile toothpicks. After all the wells are inoculated, give the plate a gentle swirl to spread the cells on the surface of the medium in each well. This makes semi-liquid cultures for use as template for colony PCR, after which the plates can be parafilmed and stored. Sending strains: 150µl per well Let plates sit on bench to dry for several days before inoculating wells. Do NOT add water to the wells. from Linda Riles, November 1997