各类藻类培养基完整资料doc.docx

各类藻类培养基完整资料doc.docx

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各类藻类培养基完整资料doc

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AJS（AcidifiedJM:

SE）

FreshwateralgaDunaliellaacidophila

MediumAcidified97:

3mixtureofJMandSE2

Seeseparaterecipes（JMandSE2）.Forapproximately1 litreoffinalmedium,mix970 mlJMwith30 mlSE2.Add10 mlofconcH2SO4togiveapHofapproximately1.5.Autoclaveat15psifor15minutes.

ANT（Antia'sMedium）

Marinealgae

StockTracemetalsstocksolution（chelated）

perlitre

EDTANa2.2H2O3.24g

FeCl3.4H2O1.08g

MnSO4.4H2O0.450g

ZnSO4.7H2O0.230g

Na2MoO4.2H2O0.097g

CuSO4.5H2O0.01g

CoSO4.7H2O0.0056g

Makeupto1litrewithdistilledwaterandadjustpHto7.6-7.8withdiluteHClorNaOH.Storefrozen.

Mediumperlitre

KNO30.05g

NaH2PO4.2H2O0.0078g

Tris[tris（hydroxymethyl）aminomethane]1.0g

Glycine0.3g

Tracemetalsstocksolution（chelated）2.5ml

ThiamineHCl500.0µg

Cyanocobalamin2.0µg

Biotin1.0µg

Filterednaturalseawater800.0ml

Makeupto1litrewithdistilledwaterandautoclaveat15psi.FinalpHshouldbe7.6-7.8.

RefAntia,Cheng&Taylor（1969）

ASW（ArtificialSeawater）

Marinealgae

Stocks

（1）Extrasaltsperlitre

NaNO330.0g

Na2HPO41.2g

K2HPO41.0g

（2）Vitaminsolutionperlitre

Biotin0.0002g

Calciumpantothenate0.02g

Cyanocobalamin0.004g

Folicacid0.0004g

Inositol1.0g

Nicotinicacid0.02g

ThiamineHCl0.1g

Thymine0.6g

Maybestoredfrozenat-20°C

Mediumperlitre

*"UltramarineSynthetica"seasalts33.6g

Extrasaltsstocksolution

（1）3.75ml

Vitaminstocksolution

（2）2.5ml

Soilextract（SE1-seerecipe）25.0ml

Tricine0.50g

Makeupto1litrewithdistilledwaterandadjustpHto7.6-7.8with1MNaOHorHCl.Autoclaveat15psi.

Supply*WaterlifeResearchIndustriesLtd,476BathRoad,Longford,WestDrayton,MiddlesexUB70ED,UK.Tel（01753）685696Fax（01753）685437

ASW+barley（ArtificialSeawater+barleygrains）

MarinedinoflagellateOxyrrhisMedium

FollowtherecipeforASWbutaddapproximatelyonegrainofbarleytoeach25mlofpreparedmediumpriortoautoclaving.

2ASW（doublestrengthArtificialSeawater）

Marinealgae

Medium

FollowtherecipeforASWbutadd35gNaClperlitrepriortoautoclaving.

ASW:

BG

Marinealgae

Medium1:

1mixture

Seeseparaterecipes.Mixaseptically.

ASW:

SES

Marinealgae

Medium3:

1mixture

Seeseparaterecipes.Mixbeforeautoclaving.

ASWP（ArtificialSeawaterforProtozoa）

Marineprotozoa

StockSolution

（1）perlitre

NaNO35.625g

Na2HPO40.225g

K2HPO40.188g

Mediumperlitre

*"UltramarineSynthetica"seasalts33.6g

Tricine0.5g

Stocksolution

（1）10.0ml

Soilextract（SE2-seerecipe）50.0ml

Makeupto1litrewithdeionizedwaterandadjustpHto7.6-7.8with1MNaOHorHCl.Autoclaveat15psifor15minutes.

Supply*asinASWrecipe

BB（Bold'sBasalMedium）

Freshwateralgae

Stocksper200ml

（1）NaNO35.0g

（2）MgSO4.7H2O1.5g

（3）NaCl0.5g

（4）K2HPO41.5g

（5）KH2PO43.5g

（6）CaCl2.2H2O0.5g

（7）Traceelementssolution:

perlitre

ZnSO4.7H2O8.82g

MnCl2.4H2O1.44g

MoO30.71g

CuSO4.5H2O1.57g

Co（NO3）2.6H2O0.49g

Mayneedautoclavingtodissolve.

per100ml

（8）H3BO31.14g

（9）EDTA-KOHsolution:

EDTANa25.0g

KOH3.1g

perlitre

（10）FeSO4.7H2O4.98g

concH2SO41.0ml

MediumStocksolutions1-610.0mleach

Stocksolutions7-101.0mleach

Makeupto1litrewithdeionizedwater.Foragar,add15.0gperlitreof*BacteriologicalAgar（OxoidL11）.Autoclaveat15psifor15minutes.

Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UK

BB:

MErds

Freshwater/brackishwateralgae

Medium8:

2mixtureor1:

1mixture

Seeseparaterecipes（BBandMErds）.Mixthenautoclaveat15psifor15minutes.

BG（Blue-GreenMedium）

Marinecyanobacteria

Stocks

（1）Extranutrientsaltsper100ml

NaNO33.00g

Na2HPO40.12g

K2HPO40.10g

（2）Tracemetalsolutionper100ml

H3BO30.286g

MnCl2.4H2O0.022g

Na2MoO4.2H2O0.039g

CuSO4.5H2O0.008g

Co（NO3）2.6H2O0.005g

MediumThismediumismadeupin2parts:

PART1

perlitre

Tricine0.50g

Soilextract（SE1-seerecipe）25.00ml

Extranutrientsalts

（1）3.75ml

Makeupto1litrewithfilterednaturalseawaterandadjustpHto7.6-7.8with1MNaOHorHCl.

PART2

perlitre

NaNO31.500g

K2HPO4.3H2O0.040g

MgSO4.7H2O0.075g

CaCl2.2H2O0.036g

Citricacid0.006g

Ammoniumferriccitrategreen0.006g

EDTANa20.001g

Na2CO30.020g

Tracemetalsolution

（2）1.00ml

Makeupto1litrewithdistilledwaterandadjustpHto7.4.

Final

AutoclaveParts1and2separatelyat15psi,allowtocoolthenmixaseptically.

Foragarplates,add15gnon-nutrientagarperlitre.

BG11（Blue-GreenMedium）

Freshwateralgaeandprotozoa

Stocksperlitre

（1）NaNO315.0g

per500ml

（2）K2HPO42.0g

（3）MgSO4.7H2O3.75g

（4）CaCl2.2H2O1.80g

（5）Citricacid0.30g

（6）Ammoniumferriccitrategreen

（枸橼酸铁铵,柠檬酸铁铵）0.30g

（7）EDTANa20.05g

（8）Na2CO31.00g

（9）Tracemetalsolutionperliter

H3BO32.86g

MnCl2.4H2O1.81g

ZnSO4.7H2O0.22g

Na2MoO4.2H2O0.39g

CuSO4.5H2O0.08g

Co（NO3）2.6H2O0.05g

MediumStocksolution1100.0ml

Stocksolutions2-810.0mleach

Stocksolution91.0ml

Makeupto1litrewithdeionizedwater.AdjustpHto7.1with1MNaOHorHCl.Foragaradd15.0gperlitreof*BacteriologicalAgar（OxoidL11）.Autoclaveat15psifor15minutes.

Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UK

RefStanieretal,（1971）

CH（Chalkley'sMedium）

Freshwaterprotozoa

Stocksper100ml

（1）NaCl2.0g

（2）KCl0.08g

（3）CaCl20.12g

MediumStocksolutions1-35.0mleach

Makeupto1litrewithdeionizedwater.Autoclaveat15psifor15minutes.

CHM（ChilomonasMedium）

Freshwaterprotozoa

Mediumperlitre

Sodiumacetatetrihydrate1.0g

*"Lab-Lemco"powder（OxoidL29）1.0g

Addtheaboveconstituentsto1litreofdeionizedwater.Autoclaveat15psifor15minutes.

Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UK

CMA（CornMealGlucoseAgar）

Freshwaterprotozoa

Mediumperlitre

*Cornmealagar（OxoidCM103）17.0g

D-glucose2.0g

*Yeastextract（OxoidL21）1.0g

Dispersetheagarincolddeionizedwater.Applyheat,stirringconstantly.Duringheatingaddglucoseandyeastextract.Bringcontentstoboil.Transfermoltenagartosuitablevessels.Sterilizebypressurecookingat10psifor15minutes.

Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UK

DM（DiatomMedium）

Freshwateralgae

Stocksper200ml

（1）Ca（NO3）2.4H2O4.00g

（2）KH2PO42.48g

（3）MgSO4.7H2O5.00g

（4）NaHCO33.18g

（5）EDTAFeNa0.45g

EDTANa20.45g

（6）H3BO30.496g

MnCl2.4H2O0.278g

（NH4）6Mo7O24.4H2O0.20g

（7）Cyanocobalamin0.008g

ThiamineHCl0.008g

Biotin0.008g

*（8）NaSiO3.9H20（SigmaS4392）11.4g

MediumStocksolutions1-81.0mleach

Makeupto1litrewithdeionizedwater.AdjusttopH6.9with1MHCl.Dispensetosuitablevesselsandautoclaveat15psifor15minutes.

Supply*SigmaChemicalCo,FancyRoad,Poole,DorsetBH177TG,UK

RefBeakes,Canter&Jaworski（l988）

EG（EuglenaGracilisMedium）

Freshwateralgaeandprotozoa

Stock

（1）CaCl2stocksolution:

perlitre

CaCl21.0g

Mediumperlitre

Sodiumacetatetrihydrate1.0g

*"Lab-Lemco"powder（OxoidL29）1.0g

*Tryptone（OxoidL42）2.0g

*Yeastextract（OxoidL21）2.0g

CaCl2stocksolution

（1）10.0ml

Addconstituentsaboveandmakeupto1litrewithdeionizedwater.Foragaradd15gperlitre*BacteriologicalAgar（OxoidL11）.Autoclaveat15psifor15minutes.

Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UK

EG:

JM

Medium1:

1mixture

Seeseparaterecipes（EGandJM）.Mixthenautoclaveat15psifor15minutes.

E27（E27Medium）

Marinealgae,includingsterilitytesting

MediumThismediumismadeupin2partstoavoidprecipitation:

PART1

perlitre

Soilextract（SE1-seerecipe）25.0ml

KNO30.050g

K2HPO40.005g

MgSO4.7H200.005g

Glucose0.250g

*Tryptone（OxoidL42）0.025g

*Liverdigest（OxoidL27）0.025g

Cyanocobalamin100.00ng

ThiamineHCl50.00µg

Makeupto500mlwithdistilledwater.

PART2

Filterednaturalseawater500.0ml

FinalAutoclaveparts1and2separatelyat15psi,allowtocoolthenmixaseptically.Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UK

E31（E31Medium）

Marinealgae

MediumThismediumismadeupin2partstoavoidprecipitation:

PART1

perlitre

Soilextract（SE1-seerecipe）50.0ml

KNO30.10g

K2HPO40.01g

MgSO4.7H200.01g

Cyanocobalamin100.00ng

ThiamineHCl50.00µg

Biotin100.0ng

Makeupto500mlwithdistilledwater.

PART2

Filterednaturalseawater500.0ml

FinalAutoclaveParts1and2separatelyat15psi,allowtocoolthenmixaseptically.

E31:

ANT

Medium1:

1mixture

Seeseparaterecipes（E31andANT）.Autoclaveseparately.Mixasepticallywhencool.