各類藻類培養(yǎng)基

1、AJS(AcidifiedJM:SE)FreshwateralgaunaliellaacidhilaMediumAcidified97:3mixtureofJMandSE2Seeseparaterecipes(and).Forapproximately1litreoffinalmedium,mix970mlJMwith30mlSE2.Add10mlofconcH2s04togiveapHofapproximately1.5.Autoclaveat15psifor15minutes.ANT(AntiasMedium)MarinealgaeStockTracemetalsstocksolution

2、EDTANa2.2H2OFeCl3.4H2OMnSO4.4H2OZnSO4.7H2ONa2MoO4.2H2OCuSO4.5H2OCoSO4.7H2O(chelated)perlitre3.24g1.08g0.450g0.230g0.097g0.01g0.0056gMakeupto1litrewithdistilledwaterandadjustpHto7.6-7.8withdiluteHClorNaOH.Storefrozen.MediumperlitreKNO30.05gNaH2PO4.2H2O0.0078gTristris(hydroxymethyl)aminomethane1.0gGly

3、cine0.3gTracemetalsstocksolution(chelated)2.5mlThiamineHCl500.0pgCyanocobalamin2.0pgBiotin1.0pgFilterednaturalseawater800.0mlMakeupto1litrewithdistilledwaterandautoclaveat15psi.FinalpHshouldbe7.6-7.8.efASW(ArtificialSeawater)MarinealgaeStocks(1)ExtrasaltsperlitreNaNO330.0gTOC o 1-5 h zNa2HPO41.2gK2H

4、PO41.0g(2)VitaminsolutionperlitreBiotin0.0002gCalciumpantothenate0.02gCyanocobalamin0.004gFolicacid0.0004gInositol1.0gNicotinicacid0.02gThiamineHCl0.1gThymine0.6gMaybestoredfrozenat-20DCMediumperlitre*UltramarineSyntheticaseasalts33.6gExtrasaltsstocksolution(1)3.75mlVitaminstocksolution(2)2.5mlSoile

5、xtract()25.0mlTricine0.50gMakeupto1litrewithdistilledwaterandadjustpHto7.6-7.8with1MNaOHorHCl.Autoclaveat15psi.Supply*WaterlifeResearchIndustriesLtd,476BathRoad,Longford,WestDrayton,MiddlesexUB70ED,UK.Tel(01753)685696Fax(01753)685437ASW+barley(ArtificialSeawater+barleygrains)Marinedinoflagellateyrrh

6、isMediuFollowtherecipeforASWbutaddapproximatelyonegrainofbarleytoeach25mlofpreparedmediumpriortoautoclaving.2ASW(doublestrengthArtificialSeawater)MarinealgaeMediumFollowtherecipeforASWbutadd35gNaClperlitrepriortoautoclaving.ASW:BGMarinealgaeMedium1:1mixtureSeeseparaterecipes.Mixaseptically.ASW:SESMa

7、rinealgaeMedium3:1mixtureSeeseparaterecipes.Mixbeforeautoclaving.ASWP(ArtificialSeawaterforProtozoa)MarineprotozoaStockSolution(1)perlitreNaNO35.625gNa2HPO40.225gK2HPO40.188gMediumperlitre*UltramarineSyntheticaseasalts33.6gTricine0.5gStocksolution(1)10.0mlSoilextract(SE2-seerecipe)50.0mlMakeupto1lit

8、rewithdeionizedwaterandadjustpHto7.6-7.8with1MNaOHorHCl.Autoclaveat15psifor15minutes.Supply*asinBB(BoldsBasalMedium)FreshwateralgaeStocksper200ml(1)NaNO35.0g(2)MgSO4.7H2O1.5g(3)NaCl0.5g(4)K2HPO41.5g(5)KH2PO43.5g(6)CaCl2.2H2O0.5gTraceelementssolution:perlitreZnSO4.7H2O8.82gMnCl2.4H2O1.44gMoO30.71gCuS

9、O4.5H2O1.57gCo(NO3)2.6H2O0.49gMayneedautoclavingtodissolve.per100ml(8)H3BO31.14g(9)EDTA-KOHsolution:EDTANa25.0gKOH3.1gperlitre(10)FeSO4.7H2O4.98gconcH2SO41.0mlMediumStocksolutions1-610.0mleachStocksolutions7-101.0mleachMakeupto1litrewithdeionizedwater.Foragar,add15.0gperlitreofBacteriologicalAgar(Ox

10、oidL11).Autoclaveat15psifor15minutes.Suppl*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKBB:MErdsFreshwater/brackishwateralgaeMedium8:2mixtureor1:1mixtureSeeseparaterecipes(and).Mixthenautoclaveat15psifor15minutes.BG(Blue-GreenMedium)MarinecyanobacteriaStocks(1)Extranutrientsaltsper100mlNaNO33.00gN

11、a2HPO40.12gK2HPO40.10g(2)Tracemetalsolutionper100mlH3BO30.286gMnCl2.4H2O0.022gTOC o 1-5 h zNa2MoO4.2H2O0.039gCuSO4.5H2O0.008gCo(NO3)2.6H2O0.005gMediumThismediumismadeupin2parts:perlitreTricine0.50gSoilextract(-seerecipe)25.00mlExtranutrientsalts(1)3.75mlMakeupto1litrewithfilterednaturalseawaterandad

12、justpHto7.6-7.8with1MNaOHorHCl.perlitreTOC o 1-5 h zNaNO31.500gK2HPO4.3H2O0.040gMgSO4.7H2O0.075gCaCl2.2H2O0.036gCitricacid0.006gAmmoniumferriccitrategreen0.006gEDTANa20.001gNa2CO30.020gTracemetalsolution(2)1.00mlMakeupto1litrewithdistilledwaterandadjustpHto7.4.FinalAutoclaveParts1and2separatelyat15p

13、si,allowtocoolthenmixaseptically.Foragarplates,add15gnon-nutrientagarperlitre.BG11(Blue-GreenMedium)FreshwateralgaeandprotozoaStocksperlitreTOC o 1-5 h zNaNO315.0gper500mlK2HPO42.0gMgSO4.7H2O3.75gCaCl2.2H2O1.80gCitricacid0.30gAmmoniumferriccitrategreen(枸櫞酸鐵銨，檸檬酸鐵銨)0.30gEDTANa20.05gNa2CO31.00g(9)Trac

14、emetalsolutionperliterH3BO32.86gMnCl2.4H2O1.81gZnSO4.7H2O0.22gNa2MoO4.2H2O0.39gCuSO4.5H2O0.08gCo(NO3)2.6H2O0.05gMediumStocksolution1100.0mlStocksolutions2-810.0mleachStocksolution91.0mlMakeupto1litrewithdeionizedwater.AdjustpHto7.1with1MNaOHorHCl.Foragaradd15.0gperlitreofBacteriologicalAgar(OxoidL11

15、).Autoclaveat15psifor15minutes.Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKRefCH(ChalkleysMedium)FreshwaterprotozoaStocksper100mlTOC o 1-5 h zNaCl2.0gKCl0.08gCaCl20.12gMediumStocksolutions1-35.0mleachMakeupto1litrewithdeionizedwater.Autoclaveat15psifor15minutes.CHM(ChilomonasMedium)Freshwa

16、terprotozoaMediumperlitreSodiumacetatetrihydrate1.0g*Lab-Lemcopowder(OxoidL29)1.0gAddtheaboveconstituentsto1litreofdeionizedwater.Autoclaveat15psifor15minutes.Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKCMA(CornMealGlucoseAgar)FreshwaterprotozoaMediumperlitre*Cornmealagar(OxoidCM103)17.0gD

17、-glucose2.0g*Yeastextract(OxoidL21)1.0gDispersetheagarincolddeionizedwater.Applyheat,stirringconstantly.Duringheatingaddglucoseandyeastextract.Bringcontentstoboil.Transfermoltenagartosuitablevessels.Sterilizebypressurecookingat10psifor15minutes.Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKD

18、M(DiatomMedium)FreshwateralgaeStocksper200ml(1)Ca(NO3)2.4H2O4.00g(2)KH2PO42.48g(3)MgSO4.7H2O5.00g(4)NaHCO33.18g(5)EDTAFeNa0.45gEDTANa20.45g(6)H3BO30.496gMnCl2.4H2O0.278g(NH4)6Mo7O24.4H2O0.20g(7)Cyanocobalamin0.008gThiamineHCl0.008gBiotin0.008g*(8)NaSiO3.9H20(SigmaS4392)11.4gMediumStocksolutions1-81.

19、0mleachMakeupto1litrewithdeionizedwater.AdjusttopH6.9with1MHCl.Dispensetosuitablevesselsandautoclaveat15psifor15minutes.Supply*SigmaChemicalCo,FancyRoad,Poole,DorsetBH177TG,UKRefEG(EuglenaGracilisMedium)Stock(1)CaCl2stocksolution:perlitre1.0gFreshwateralgaeandprotozoaStock(1)CaCl2stocksolution:perli

20、tre1.0gCaCl2MediumperlitreSodiumacetatetrihydrate1.0g*Lab-Lemcopowder(OxoidL29)1.0g*Tryptone(OxoidL42)2.0g*Yeastextract(OxoidL21)2.0gCaCl2stocksolution(1)10.(0mlAddconstituentsaboveandmakeupto1litrewithdeionizedwater.Foragaradd15gperlitreBacteriologicalAgar(OxoidL11).Autoclaveat15psifor15minutes.Sup

21、ply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKEG:JMMedium1:1mixtureSeeseparaterecipes(and).Mixthenautoclaveat15psifor15minutes.E27(E27Medium)Marinealgae,includingsterilitytestingMediumThismediumismadeupin2partstoavoidprecipitation:PART1perlitreSoilextract()25.0mlKNO30.050gK2HPO40.005gMgSO4.7H200.005gGlucose0.250g*Tryptone(OxoidL42)0.025g*Liverdigest(OxoidL27)0.025gCyanocobalamin100.00ngThiamineHCl5