各類(lèi)藻類(lèi)培養(yǎng)基(完整資料).doc

1、【最新整理，下載后即可編輯】AJS (Acidified JM:SE)Freshwater alga Dun疝ella addophilaMedium Acidified 97:3 mixture of JM and SE2Sec separate rccipesQM and SE2). For approximately 1 litre of final medium, mix 970 ml JM with 30 ml SE2. Add 10 ml of cone H2s(% to give a pH of approximately 1.5. Autoclave at 15 psi for

2、 15 minutes.ANT (Antia's Medium)Marine algaeStock Trace metals stock solution (chelated)per litreEDTANa2.2H2O3.24 gFeC13.4H2O1.08gMnSO4.4H2O0.450 gZnSO4.7H2O0.230 gNa2MoO4.2H2O0.097 gCuS()4.5H2O0.01gCoSO4.7H2O0.0056 gNaN()330.0 g1.2 g1.() gNa2HP()4K2HPO4(2) Vitamin solution per litreBiotin0.0002

3、 gCalciumpantothenate0.02 gCvanocobalamin J0.004 gFolicacid().0()04 gInositol1.() gNicotinicacid0.02 gThiamineHClo.igThymine J0.6 gMay be stored frozen at -20° CMediumper litreUltramarine Synthetica11seasalts33.6 gExtrasaltsstocksolution【最新整理，下載后即可編輯】3.75 mlVitamin stock solution (2) 2.5 mlSoil

4、 extract (SEI - see recipe) 25.0 mlTricine0.50 gMake up to 1 litre with distilled water and adjust pH to 7.6 - 7.8 with IM NaOH or HC1. Autoclave at 15 psi.Supply * Waterlife Research Industries Ltd, 476 Bath Road, Longford, West Drayton, Middlesex UB7 OED, UK. Tel (01753) 685696 Fax (01753) 685437A

5、SW + barley (Artificial Seawater + barley grains)Marine dinoflagellate Oxyrrhis MediumFollow the recipe for ASW but add approximately one grain of barley to each 25 ml of prepared medium prior to autoclaving.2ASW (double strength Artificial Seawatei)Marine algaeMediumFollow the recipe for ASW but ad

6、d 35 g NaCl per litre prior to autoclaving.ASW:BGMarine algae【最新整理，下載后即可編輯】Medium 1:1 mixtureSee separate recipes. Mix aseptically.ASW:SESMarine algaeMedium 3:1 mixtureSee separate recipes. Mix before autoclaving.ASWP (Artificial Seawater for Protozoa)Marine protozoaStockSolution(1)per litre NaNO35.

7、625 gNa2HP()40.225 gK2HPO40.188 gMedium per litre *HUltramarinc SvnthcticaM sea saltsJ33.6 gTricinc0.5 gStocksolution(1)10.0 mlSoil extract (SE2 - see recipe)【最新整理，下載后即可編輯】50.0 mlMake up to 1 litre with deionized water and adjust pH to 7.6 - 7.8 with IM NaOH or HC1. Autoclave at 15 psi for 15 minute

8、s. Supply * as in ASW recipeBB (Bold's Basal Medium)Freshwater algaeStocks per 200 ml(1) NaN()35.Og(2) MgSO4.7H2O1.5 gNaCl0.5 g(4) K2HPO4L5g(5) KH2PO43.5 g(6) CaC12.2H2O0.5 g(7) Trace elements solution:per litreZnSO4.7H2O8.82 gMnC12.4H2O1.44 gMoO3【最新整理，下載后即可編輯】0.71gCuSO4.5H2O1.57 gCo(NO3)2.6H2O0

9、.49 gMay need autoclaving to dissolve.per 100 ml(8) H3BQ31.14g(9) EDTA - KOH solution: EDTANa25.OgKOH9.1 gper litre(10) FcSO4.7H2O4.98 gconeH2SO41.0 mlMediumStock solutions 1-610.0 ml eachStock solutions 7-101.() ml eachMake up to 1 litre with deionized water. For agar, add 15.() g per litre of *Bac

10、teriological Agar (Oxoid LI 1). Autoclave at 15 psi for 15 minutes.Supply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24OPW, UKBB:MErdsFreshwater/brackish water algaeMedium 8:2 mixture or 1:1 mixtureSec separate recipes (BB and MErds). Mix then autoclave at 15 psi for 15 minutes.BG (Blue-Green Me

11、dium)Marine cyanobacteriaStocks (1)Extra nutrient saltsper 100 mlNaNO33.00 gNa2HPO40.12 gK2HPO40.10 g(2) per 100 mlTrace metal solutionH3BO30.286 gMnC12.4H2O0.022 gNa2MoO4.2H2O0.039 gCuSO4.5H2O0.008 gCo(N()3)2.6H2()0.005 gMedium This medium is made up in 2 parts:PARTIper litreTricine0.50 gSoil extra

12、ct (SEI - see recipe) 25.00 mlExtranutrientsalts(1)3.75 mlMake up to 1 litre with filtered natural seawater and adjust pH to 7.6 - 7.8 with IM NaOH or HC1.PART 2per litreNaNO31.500 gK2HPO4.3H2O0.040 gMgSO4.7H2O0.075 gCaC12.2H2O0.036 gCitricacid0.006 gAmmonium ferric citrate green0.006 gEDTANa20.001

13、gNa2co30.02() gTracemetalsolution(2)1.00 mlMake up to 1 litre with distilled water and adjust pH to 7.4.FinalAutoclave Parts 1 and 2 separately at 15 psi, allow to cool then mix aseptically.For agar plates, add 15 g non-nutrient agar per litre.【最新整理，下載后即可編輯】BG11 (Blue-Green Medium)Freshwater algae a

14、nd protozoaStocks per litre 15.0 gper 500 ml(2)2.0 g(3)3.75 g(4)1.8() g 0.30 gNaN()3K2HPO4MgS()4.7H2OCaC12.2H2OCitricacid(6) Ammonium ferric citrate green(枸椽酸鐵錠，檸檬酸鐵錢(qián))().3() gEDTANa20.05 g(8)1.00g(9)Traceper literH3BO32.86 gMnC12.4H2O1.81gZnSO4.7H2O0.22 gNa2Mo()4.2H2()().39 gCuS()4.5H2()0.08 gCo(N()

15、3)2.6H2()0.05 gMediumStock100.0 mlmetalsolutionStocksolutions(7)Na2c()3solution10.() ml eachStocksolution1.() mlMake up to 1 litre with deionized water. Adjust pH to 7.1 withIM NaOH or HO. For agar add 15.() g per litre of"Bacteriological Agar (Oxoid LI 1). Autoclave at 15 psi for 15 minutes.Su

16、pply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24 OPW, UK7?e/'Stanier et al, (1971)CH (Chalkleyfs Medium)Freshwater protozoaStocks per 100 ml(1) NaCl2.0 g(2) KC10.08 g(3) CaC120.12 gMediumStock solutions 1-35.() ml eachMake up to 1 litre with deionized water. Autoclave at 15 psi for 15 minu

17、tes.CHM (Chilomonas Medium)Freshwater protozoaMediumper litreSodiumacetatetrihvdrateJ1.() g*nLab-LemcoH powder (Oxoid L29) 1.0 g Add the above constituents to 1 litre of deionized water. Autoclave at 15 psi for 15 minutes.Supply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24 OPW, UKCMA (Com Meal

18、Glucose Agar)Freshwater protozoaMediumper litre*Corn meal agar (Oxoid CM 103)".OgD-glucose2.0 g*Yeastextract(OxoidL21)1.() gDisperse the agar in cold deionized water. Apply heat, stirring constantly. During heating add glucose and yeast extract. Bring contents to boil. Transfer molten agar to s

19、uitable vessels. Sterilize by pressure cooking at 10 psi for 15 minutes.Supply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24 0PW, UKDM (Diatom Medium)Freshwater algaeStocks per 200 mlCa(NO3)2.4H2O4.00 g2.48 g(2)KH2PO45.00 g(3)MgS()4.7H2()3.18 g(4)NaHCO30.45 g().45 g(5)EDTANa2EDTAFeNa0.496 g0.278

20、 g0.20 g(6)MnC12.4H2O(NH4)6Mo7()24.4H2()H3BO30.008 gCvanocobalamin J0.008 g0.008 gThiamineBiotinHC111.4g*(8)NaSiO3.9H20(Sigma S4392)MediumStock solutions 1-81.() ml eachMake up to 1 litre with deionized water. Adjust to pH 6.9 with IM HC1. Dispense to suitable vessels and autoclave at 15 psi for15 m

21、inutes.Supply * Sigma Chemical Co, Fancy Road, Poole, Dorset BH17 7TG, UK7?c/'Bcakes, Canter & Jaworski (1988)EG (Eugjena Gracilis Medium)Freshwater algae and protozoaStockCaC12stocksolution:per litreCaC121.0 gMediumper litreSodiumacetatetrihydratcJ1.0 g*HLab-LcmcoHpowder(OxoidL29)1.0 g*Tryp

22、tone(OxoidL42)2.0 g*Yeastextract(OxoidL21)2.0 gCaC12stocksolution10.0 mlAdd constituents above and make up to 1 litre with deionized water. For agar add 15 g per litre *Bacteriological Agar (Oxoid LI 1). Autoclave at 15 psi for 15 minutes.Supply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24 0PW,

23、 UKEG:JMMedium 1:1 mixtureSee separate recipes (EG and JM). Mix then autoclave at 15psi for 15 minutes.E27 (E27 Medium)Marine algae, including sterility testingMedium This medium is made up in 2 parts to avoid precipitation:PARTIper litreSoil extract (SEI - see recipe) 25.0 mlKNO30.050 gK2HPO40.005 gMgSO4.7H2()0.005 gGlucose0.25() g*Tryptonc(OxoidL42)0.025 g* Liverdigest(OxoidL27)0.025 g Cyanocobal