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

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