各類藻類培養(yǎng)基

1、文檔來源為：從網(wǎng)絡(luò)收集整理.word版本可編輯.歡迎下載支持AJS (Acidified JM:SE)Freshwater alga Dunaliella acidophilaMedium Acidified 97:3 mixture of JM and SE2See separate recipes( JM and SE2 ). For approximately 1 litre of final medium, mix 970 ml JM with 30 ml SE2. Add 10 ml of conc H2SO 4 to give a pH of approximately 1.5.

2、Autoclave at 15 psifor 15 minutes.ANT (Antia's Medium)Marine algaeStock Trace metals stock solution (chelated) per litre EDTANa2.2H2O3.24gFeCl3.4H2O1.08 gMnSO4.4H2O0.450gZnSO4.7H2O0.230gNa2MoO4.2H2O0.097gCuSO4.5H2O0.01 gCoSO4.7H2O0.0056gMake up to 1 litre with distilled water and adjust pH to 7.

3、6 - 7.8 with dilute HCl or NaOH. Store frozen.Mediumper litreKNO30.05 gNaH2PO4.2H2O0.0078 gTris tris(hydroxymethyl)aminomethane 1.0 gGlycine0.3 gTrace metals stock solution (chelated) 2.5 mlThiamine HCl500.0聞Cyanocobalamin2.0聞Biotin1.0聞Filtered natural seawater800.0 mlMake up to 1 litre with distill

4、ed water and autoclave at 15 psi. Final pH should be 7.6 - 7.8.Ref Antia, Cheng & Taylor (1969)ASW (Artificial Seawater)Marine algaeStocks (1) Extra saltsper litre30.0 g1.2 g1.0 gNaNO3Na2HPO4K2HPO4文檔來源為：從網(wǎng)絡(luò)收集整理.word版本可編輯.歡迎下載支持(2) Vitamin solution per litreBiotin0.0002 gCalcium pantothenate0.02

5、gCyanocobalamin0.004 gFolic acid0.0004 gInositol1.0 gNicotinic acid0.02 gThiamine HCl0.1 gThymine0.6 gMay be stored frozen at -20° CMediumper litre*"Ultramarine Synthetica" sea salts 33.6 gExtra salts stock solution (1)3.75 mlVitamin stock solution (2)2.5 mlSoil extract (SE1 - see rec

6、ipe )25.0 mlTricine0.50 gMake up to 1 litre with distilled water and adjust pH to 7.6 - 7.8 with 1M NaOH or HCl. Autoclave at 15 psi.Supply * Waterlife Research Industries Ltd, 476 Bath Road, Longford, West Drayton, Middlesex UB7 0ED, UK. Tel (01753) 685696 Fax (01753) 685437ASW + barley (Artificial

7、 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 Seawater)Marine algaeMediumFollow the recipe for ASW but add 35 g NaCl per litre p

8、rior to autoclaving.ASW:BGMarine algaeMedium 1:1 mixtureSee separate recipes. Mix aseptically.ASW:SES文檔來源為：從網(wǎng)絡(luò)收集整理.word版本可編輯.歡迎下載支持Marine algaeMedium 3:1 mixtureSee separate recipes. Mix before autoclaving.ASWP (Artificial Seawater for Protozoa)Marine protozoaStock Solution (1)NaNO3per litre5.625 gN

9、a2HPO4K2HPO40.225 g0.188 gMediumper litre*"Ultramarine Synthetica" sea saltsTricine0.533.6 gStock solution (1)Soil extract (SE2 - see recipe)10.0 ml50.0 mlMake up to 1 litre with deionized waterand adjust pH to 7.6 - 7.8 with 1M NaOH or HCl.Autoclave at 15 psi for 15 minutes.Supply * as in

10、 ASW recipeBB (Bold's Basal Medium)Stocksper 200 ml(1) NaNO35.0 g MgSO4.7H2O1.5 g(3) NaCl0.5 g(4) K2HPO41.5 g KH2PO43.5 g(6) CaCl2.2H2O0.5 g(7) Trace elements solution:per litreZnSO4.7H2O8.82 gMnCl2.4H2O1.44 gMoO30.71 gCuSO4.5H2O1.57 gCo(NO3)2.6H2O0.49 gMay need autoclaving to dissolve.per 100 m

11、l(8) H3BO31.14 g(9) EDTA - KOH solution:EDTANa25.0 gKOH3.1 gFreshwater algaeper litre4.98 g1.0 ml文檔來源為：從網(wǎng)絡(luò)收集整理.word版本可編輯.歡迎下載支持(10) FeSO4.7H2O conc H2SO4Medium Stock solutions 1 - 610.0 ml eachStock solutions 7 - 101.0 ml eachMake up to 1 litre with deionized water. For agar, add 15.0 g per litre of

12、 "Bacteriological Agar (Oxoid L11). Autoclave at 15 psi for 15 minutes.Supply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24 0PW, UKBB:MErdsFreshwater/brackish water algaeMedium 8:2 mixture or 1:1 mixtureSee separate recipes ( BB and MErds ). Mix then autoclave at 15 psi for 15 minutes.BG (B

13、lue-Green Medium)Marine cyanobacteriaStocks (1) Extra nutrient saltsper 100 mlNaNO33.00 gNa2HPO40.12gK2HPO40.10g(2) Trace metal solutionper 100 mlH3BO30.286gMnCl2.4H2O0.022 gNa2MoO4.2H2O0.039 gCuSO4.5H2O0.008 gCo(NO3)2.6H2O0.005 gMedium This medium is made up in 2 parts:PART 1per litreTricine0.50 gS

14、oil extract (SE1 - see recipe)25.00 mlExtra nutrient salts (1)3.75 mlMake up to 1 litre with filtered natural seawater and adjust pH to 7.6 - 7.8 with 1M NaOH or HCl.PART 2per litreNaNO31.500 gK2HPO4.3H2O0.040 gMgSO4.7H2O0.075 gCaCl2.2H2O0.036 g0.006 gCitric acid0.006 gAmmonium ferric citrate green文

15、檔來源為：從網(wǎng)絡(luò)收集整理.word版本可編輯.歡迎下載支持EDTANa20.001 gNa2CO30.020 gTrace metal solution (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.BG11 (Blue-

16、Green Medium)Freshwater algae and protozoaStocksperlitre(1) NaNO315.0 gper 500 ml(2) K2HPO42.0 g(3) MgSO4.7H2O3.75 g(4) CaCl2.2H2O1.80 g(5) Citric acid0.30 g(6) Ammonium ferric citrate green(枸檬酸鐵鏤，檸檬酸鐵鏤)0.30 g(7) EDTANa20.05 g(8) Na2CO31.00 g(9) Trace metal solutionper literH3BO32.86 gMnCl2.4H2O1.81

17、 gZnSO4.7H2O0.22 gNa2MoO4.2H2O0.39 gCuSO4.5H2O0.08 gCo(NO3)2.6H2O0.05 gMediumStock solution 1100.0 mlStock solutions 2 - 810.0 ml eachStock solution 91.0 mlMake up to 1 litre with deionized water. Adjust pH to 7.1 with 1M NaOH or HCl. For agar add 15.0 g per litre of *Bacteriological Agar (Oxoid L11

18、). Autoclave at 15 psi for 15 minutes.Supply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24 0PW, UKRef Stanier et al, (1971)CH (Chalkley's Medium)Freshwater protozoaStocks(1) NaCl(2) KClper 100 ml2.0 g0.08 g文檔來源為：從網(wǎng)絡(luò)收集整理.word版本可編輯.歡迎下載支持(3) CaCl20.12 gMedium Stock solutions 1 - 35.0 ml eachMa

19、ke up to 1 litre with deionized water. Autoclave at 15 psi for 15 minutes.CHM (Chilomonas Medium)Freshwater protozoaMediumper litreSodium acetate trihydrate1.0 g* "Lab-Lemco" powder (Oxoid L29)1.0 gAdd the above constituents to 1 litre of deionized water. Autoclave at 15 psi for 15 minutes

20、.Supply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24 0PW, UKCMA (Corn Meal Glucose Agar)Freshwater protozoaMediumper litre* Corn meal agar (Oxoid CM103)17.0 gD-glucose2.0 g* Yeast extract (Oxoid L21)1.0 gDisperse the agar in cold deionized water. Apply heat, stirring constantly. During heating

21、add glucose and yeast extract. Bring contents to boil. Transfer molten agar to suitable vessels.Sterilize by pressure cooking at 10 psi for 15 minutes.Supply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24 0PW, UKDM (Diatom Medium)Freshwater algaeStocksper 200 ml(1) Ca(NO3)2.4H2O4.00 g(2) KH2PO42.

22、48g(3) MgSO4.7H2O5.00 g(4) NaHCO33.18g(5) EDTAFeNa0.45 gEDTANa20.45 g(6) H3BO30.496gMnCl2.4H2O0.278 g(NH4)6Mo7O24.4H2O0.20 g Cyanocobalamin0.008 gThiamine HCl0.008 gBiotin0.008 g*(8) NaSiO3.9H20 (Sigma S4392)11.4 g文檔來源為：從網(wǎng)絡(luò)收集整理.word版本可編輯.歡迎下載支持Medium Stock solutions 1 - 81.0 ml eachMake up to 1 litr

23、e with deionized water. Adjust to pH 6.9 with 1M HCl. Dispense to suitablevessels and autoclave at 15 psi for 15 minutes.Supply * Sigma Chemical Co, Fancy Road, Poole, Dorset BH17 7TG, UK Ref Beakes, Canter & Jaworski (l988)EG (Euglena Gracilis Medium)Freshwater algae and protozoaStock (1) CaCl2

24、 stock solution:CaCl2per litreMedium1.0 gper litreSodium acetate trihydrate*"Lab-Lemco" powder (Oxoid L29)1.0 g1.0 g*Tryptone (Oxoid L42)*Yeast extract (Oxoid L21)CaCl2 stock solution (1)2.0 g2.0 g10.0 mlAdd constituents above and make up to 1 litre with deionized water. For agar add 15 g

25、per litre*Bacteriological Agar (Oxoid L11). Autoclave at 15 psi for 15 minutes.Supply * Unipath Ltd, Wade Road, Basingstoke, Hants RG24 0PW, 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:PART 1Soil extract (per litreSE1 - see recipe )25.0 mlKNO30.050 gK2HPO4MgSO4.7H200.005 g0.005 gGlucose0.250 g*Tryptone (Oxoid L42