yeast mating in 24-well plates
modified Clontech protocol #PT3024-1 IX/C for microtiter plates
- add 500 µl 1x YPDA (2x YPDA also works) to each well
- add 20 µl prey overnight culture
- add 20 µl bait overnight culture
or
- pick one colony of the bait and one colony of the prey and stir them into the YPDA
- place plate on a rotating platform shaker
- incubate @ 30 °C @ 120 rpm for 6 to 20 hrs
- plate 100 to 200 µl of the mating culture on a 8 cm plasmid selective plate
or
- if you know the mating works, plate 20 to 50 µl on ¼ of an 8 cm plasmid selective plate
- incubate for 2 to 5 days @ 30 °C to allow diploid cells to form colonies
- streak over and score on selective media
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