Quint Lab:yeast mating in 24-well plates

yeast mating in 24-well plates

modified Clontech protocol #PT3024-1 IX/C for microtiter plates

1. add 500 µl 1x YPDA (2x YPDA also works) to each well
2. add 20 µl prey overnight culture
3. add 20 µl bait overnight culture

or

1. pick one colony of the bait and one colony of the prey and stir them into the YPDA
2. place plate on a rotating platform shaker
3. incubate @ 30 °C @ 120 rpm for 6 to 20 hrs
4. plate 100 to 200 µl of the mating culture on a 8 cm plasmid selective plate

or

1. if you know the mating works, plate 20 to 50 µl on ¼ of an 8 cm plasmid selective plate
2. incubate for 2 to 5 days @ 30 °C to allow diploid cells to form colonies
3. streak over and score on selective media