5氟乳清酸培养基配制.pdf

琼脂培养基先灭菌冷却到50-60度时再加入用0.2um滤膜过滤除菌的5-氟乳清酸，提醒一

下，5-氟乳清酸再水里，甲、乙醇都很难溶解，在水里溶解要加热到50度左右搅拌一个小

时左右才充分溶解，溶解后用0.2um滤膜过滤除菌。我把我用的protocol给您贴出来吧！

Goodluckwithyourexperiments!

5-FOA(5-FluorooroticAcid)MediumandAgarPlateRecipe

Followingrecipemakes1liter5-FOAplates:

Step-1:Autoclave20gramsofagarin750mldoubledistilledwater.

Step-2:Addfollowingsintoaglassbeaker,andbringto250mlwithdoubledistilledwater.

AmmoniumSulfate5.0g

Yeastnitrogenbasewithoutaminoacid1.7g

Dextrose20g

Necessarynutritionalsupplements(seenote3)Xg

5-FOA1.0g

Warmup5-FOAmixtureinStep-2at55-65°Conaheatplate,stiruntilitdissolves~30min-1h.

Thesolubilityof5-FOAisverylowinwater;therefore,itisnecessarytowarmupthesolution.

toautoclaveitmyself,butyoucantryit.

After5-FOAcontainingmixtureinStep-2iscompletelydissolved,sterilizeitbyfilter.Placeboth

meltedagarand5-FOAcontainingsolutioninawaterbath(50-60°C)foronehour.Finally,add

5-FOAsolutionontomeltedagarandpouryourplates.PlatesmaybedriedatRTfor48hoursand

maybekeptat4°Cforlongtermstorage.

Notes:

muchUracil!).

2.NeveradjustthepHof5-FOAcontainingmedium.Onlyprotonatedformof5-FOAispermeable.

pHofyourplateswillbeveryacidicafteradding5-FOA.

3.Donotaddmorenutritionalsupplementsthanitisnecessary;thisreducesefficacyof5-FOA

plates.Forexample,youonlyneedtoaddHis,Leu,LysandUrasupplementsforstrainBY4742.

Thesesupplementsmaybeaddeddirectlyfrom100xstocksolution.

4.5-FOAisveryexpensivereagent.IfoundthatUSBiologicalhasthecheapestpricefor5-FOA.

5.5-FOAishighlymutagenictoCandidaalbicans.Thismayalsobethecaseforbuddingyeast.I

suggestyoudonotleaveyeastcolonieson5-FOAplatesforlongerthanitisnecessary.

WellingtonandRu