Changes

*Pick one colony and make a 1/2 inch x 1/2 inch square patch of yeaston selective plates, allow to grow for 1 day. (Does this have to be selective plates? or are YPD plates always used?24°C) *Place nitrocellulose filters on gel and incubate overnight at 23(So you're not actually moving the patch to the filter, but just allowing it to touch?)23°C. *Wash filter in water (by adding water to dish and rocking?). *Primary Wash in primary antibody (polyclonal against cpy location?) how long?overnight at 4C or >3h at room temp*Rinse in Tbst.*Secondary antibody (where/what is it and how long?)at 10 000X for 45min-1h. Wash with pbs *Rinse in between? TBST.  What is ECL?*Develop