其他

Yeast Plates

HahnLab

LastmodifiedTue,Oct6,1998

AminoAcidMix(40X)Use0.6g/literTyrosine2gArginine4gSerine2gValine2gThreonine4gIsoleucine2gPhenylalanine2gAsparticAcid2gProline2g


GlucoseCompletePlates(1liter):

1.7gYeastNitrogenbase-aminoacidsandammoniumsulfate5.0gAmmoniumSulfate1literH2OpHto7.0usingNaOHAdd:0.6gAminoAcidmix(mixwellbeforeadding)20gBacto-agar(Difco)Autoclave25min.Add:10ml(10/mg/ml)ofanyotherrequiredaminoacids10ml0.4%Adeninesulfatein0.1MHClifrequired10ml0.2%Uracilifrequired50ml40%glucose


YPDPlates(1liter):

YeastExtract10gBactopeptone20gBacto-Agar20gH2O1literAutoclave25minAdd:50ml40%glucose5ml0.4%Adeninesulfatein0.1MHClifrequiredAdd10ml0.2%Uracil(onlyifplateswillbeusedfortetraddissection)


5-FOAplates(0.5liter):

Agar:

15gBacto-agar300mlH20in500mlflaskAutoclave25min.

FOAmix:

0.85gYeastNitrogenbase-amm.sulfateandaminoacids2.5gAmmoniumsulfate0.3gaminoacidmix(fromabove)25ml40%glucose17.5mgUracil0.5g5-FOAAnyotherrequiredaminoacids225mlH2OMicrowavetheabovemixtureforabout3minuntilabout65degrees.Stironstirplateuntildissolved(~10-15min).Filterthrough0.2micronbottletopfilterintosterilebottle.Addtoautoclavedagar.Mixwellandpourplates.DonotpHtheplatesasthiswillinactivatethe5-FOA.

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