Handbook of Microbiological Media, Fourth Edition part 122 pot

20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L.. 3.0mg Preparation of Solution D: Add components to distilled/deionized water and

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Solution G 10.0mL

Solution B 1.0mL

Solution C 1.0mL

Solution D 1.0mL

pH 7.3 ± 0.2 at 25°C

Solution A:

Compositionper 940.0mL:

NaCl 1.0g

KCl 0.5g

MgCl2·6H2O 0.4g

NH4Cl 0.25g

KH2PO4 0.2g

CaCl2·2H2O 0.15g

Resazurin 0.5mg

Preparation of Solution A: Prepare under 80% N2 + 20% CO2 gas

atmosphere Add components to distilled/deionized water and bring

vol-ume to 940.0mL Mix thoroughly Adjust pH to 7.2 Sparge with 80% N2

+ 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C Cool to

25°C

Solution B:

Compositionper liter:

FeCl2·4H2O 1.5g

CoCl2·6H2O 190.0mg

MnCl2·4H2O 100.0mg

ZnCl2 70.0mg

Na2MoO4·2H2O 36.0mg

NiCl2·6H2O 24.0mg

H3BO3 6.0mg

CuCl2·2H2O 2.0mg

HCl (25% solution) 10.0mL

Preparation of Solution B: Add FeCl2·4H2O to 10.0mL of HCl

so-lution Mix thoroughly Add distilled/deionized water and bring

vol-ume to 1.0L Add remaining components Mix thoroughly Sparge with

80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C

Cool to 25°C

Solution C:

Pyridoxine hydrochloride 300.0mg

Thiamine-HCl·2H2O 200.0mg

Nicotinic acid 200.0mg

Vitamin B12 100.0mg

Calcium pantothenate 100.0mg

p-Aminobenzoic acid 80.0mg

D(+)-Biotin 20.0mg

Preparation of Solution C: Add components to distilled/deionized

water and bring volume to 1.0L Sparge with 100% N2 Mix

thorough-ly Filter sterilize

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

Preparation of Solution D: Add components to distilled/deionized

water and bring volume to 1.0L Mix thoroughly Sparge with 100%

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Preparation of Solution E: Add NaHCO3 to distilled/deionized water and bring volume to 100.0mL Mix thoroughly Sparge with 100% N2 gas mixture Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C

Solution F:

Fructose 2.0g

Preparation of Solution F: Add fructose to distilled/deionized wa-ter and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 gas mixture Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C

Solution G : Compositionper 10.0mL:

Na2S·9H2O 0.125g

Preparation of Solution G: Add Na2S·9H2O to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C Cool to 25°C

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL solu-tion F, and 10.0mL solusolu-tion G to 940.0mL solusolu-tion A Distribute anaer-obically under 80% N2 + 20% CO2 into appropriate vessels The pH should be 7.2–7.4

Use: For the cultivation of Clostridium homopropionicum DSM 5847.

Modified FWM Medium (DSMZ Medium 503)

Composition per 1013.0mL:

Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL

pH 7.3 ± 0.2 at 25°C

Solution A:

NaCl 1.0g KCl 0.5g MgCl2·6H2O 0.4g

NH4Cl 0.25g

KH2PO4 0.2g CaCl2·2H2O 0.15g Resazurin 0.5mg

Preparation of Solution A: Prepare under 80% N2 + 20% CO2 gas atmosphere Add components to distilled/deionized water and bring vol-ume to 940.0mL Mix thoroughly Adjust pH to 7.2 Sparge with 80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C

Solution B:

FeCl2·4H2O 1.5g CoCl2·6H2O 190.0mg MnCl2·4H2O 100.0mg ZnCl2 70.0mg

Na2MoO4·2H2O 36.0mg NiCl2·6H2O 24.0mg

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H3BO3 6.0mg

CuCl2·2H2O 2.0mg

Cool to 25°C

Solution C:

Vitamin B12 100.0mg

D(+)-Biotin 20.0mg

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Preparation of Solution E: Add NaHCO3 to distilled/deionized

water and bring volume to 100.0mL Mix thoroughly Sparge with

100% N2 gas mixture Autoclave for 15 min at 15 psi pressure–121°C

Cool to 25°C

Solution F:

Na2-succinate 2.5g

Preparation of Solution F: Add Na2-succinate to

distilled/deion-ized water and bring volume to 10.0mL Mix thoroughly Sparge with

Cool to 25°C

Solution G :

Na2S·9H2O 0.125g

Preparation of Solution G: Add Na2S·9H2O to distilled/deionized

water and bring volume to 10.0mL Mix thoroughly Autoclave under

100% N2 for 15 min at 15 psi pressure–121°C Cool to 25°C

N2 + 20% CO2 gas atmosphere Sequentially add 1.0mL solution B,

1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL

so-lution F, and 10.0mL soso-lution G to 940.0mL soso-lution A Distribute

an-aerobically under 80% N2 + 20% CO2 into appropriate vessels The

pH should be 7.2–7.4

Use: For the cultivation of unidentified bacterium DSM 5849

pH 7.3 ± 0.2 at 25°C

Solution A:

NH4Cl 0.25g

Solution B:

H3BO3 6.0mg CuCl2·2H2O 2.0mg HCl (25% solution) 10.0mL

Preparation of Solution B: Add FeCl2·4H2O to 10.0mL of HCl so-lution Mix thoroughly Add distilled/deionized water and bring vol-ume to 1.0L Add remaining components Mix thoroughly Sparge with 80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C

Solution C:

Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H2O 200.0mg Nicotinic acid 200.0mg Vitamin B12 100.0mg Calcium pantothenate 100.0mg

D(+)-Biotin 20.0mg

Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L Sparge with 100% N2 Mix

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

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N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Cool to 25°C

Solution F:

Na-(D/L)-3-hydroxybutyrate 1.5g

Preparation of Solution F: Add Na-(D/L)-3-hydroxybutyrate to

distilled/deionized water and bring volume to 10.0mL Mix

thorough-ly Sparge with 100% N2 gas mixture Autoclave for 15 min at 15 psi

pressure–121°C Cool to 25°C

Solution G :

Na2S·9H2O 0.125g

Use: For the cultivation of Desulfococcus biacutus DSM 5651

Solution A 940.0mL

Solution E 50.0mL

Solution F 10.0mL

Solution G 10.0mL

Solution B 1.0mL

Solution C 1.0mL

Solution D 1.0mL

pH 7.3 ± 0.2 at 25°C

Solution A:

NaCl 1.0g

KCl 0.5g

MgCl2·6H2O 0.4g

NH4Cl 0.25g

KH2PO4 0.2g

CaCl2·2H2O 0.15g

Resazurin 0.5mg

25°C

Solution B:

Solution C:

D(+)-Biotin 20.0mg

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

Preparation of Solution D: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Sparge with 100%

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Solution F:

Xylan 2.0g

Preparation of Solution F: Add xylan to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 gas mixture Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C

Na2S·9H2O 0.125g

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Use: For the cultivation of Cytophaga xylanolytica DSM 6779

Solution A 940.0mL

Solution E 50.0mL

Solution F 10.0mL

Solution G 10.0mL

Solution B 1.0mL

Solution C 1.0mL

Solution D 1.0mL

pH 7.3 ± 0.2 at 25°C

Solution A:

NaCl 1.0g

KCl 0.5g

MgCl2·6H2O 0.4g

NH4Cl 0.25g

KH2PO4 0.2g

CaCl2·2H2O 0.15g

Resazurin 0.5mg

25°C

Solution B:

FeCl2·4H2O 1.5g

CoCl2·6H2O 190.0mg

MnCl2·4H2O 100.0mg

ZnCl2 70.0mg

NiCl2·6H2O 24.0mg

H3BO3 6.0mg

CuCl2·2H2O 2.0mg

Cool to 25°C

Solution C:

Vitamin B12 100.0mg

D(+)-Biotin 20.0mg

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Solution F:

Xylose 2.0g

Preparation of Solution F: Add xylose to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 gas mixture Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C

Na2S·9H2O 0.125g

N2 + 20% CO2 gas atmosphere Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so-lution F, and 10.0mL soso-lution G to 940.0mL soso-lution A Distribute an-aerobically under 80% N2 + 20% CO2 into appropriate vessels The

Use: For the cultivation of Cytophaga xylanolytica DSM 6779

pH 7.3 ± 0.2 at 25°C

Solution A:

NH4Cl 0.25g

KH2PO4 0.2g

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CaCl2·2H2O 0.15g

Resazurin 0.5mg

25°C

Solution B:

FeCl2·4H2O 1.5g

CoCl2·6H2O 190.0mg

MnCl2·4H2O 100.0mg

ZnCl2 70.0mg

NiCl2·6H2O 24.0mg

H3BO3 6.0mg

CuCl2·2H2O 2.0mg

Cool to 25°C

Solution C:

Vitamin B12 100.0mg

D(+)-Biotin 20.0mg

water and bring volume to 1.0L Sparge with 100% N2 Mix thoroughly

Filter sterilize

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Cool to 25°C

Solution F:

Pivalic acid 1.0g

NaNO3 0.85g

Preparation of Solution F: Add components to distilled/deionized

water and bring volume to 10.0mL Mix thoroughly Sparge with 100%

N2 gas mixture Autoclave for 15 min at 15 psi pressure–121°C Cool

to 25°C

Na2S·9H2O 0.125g

Use: For the cultivation of Thavera pivalivorans DSM 14691

pH 7.3 ± 0.2 at 25°C

Solution A:

NH4Cl 0.25g

Solution B:

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Solution C:

Vitamin B12 100.0mg

D(+)-Biotin 20.0mg

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Cool to 25°C

Solution F:

Glucose 5.0g

Yeast extract 2.0g

to 25°C

Solution G :

Na2S·9H2O 0.125g

Use: For the cultivation of Opitutus sp DSM 14424.

Solution A 940.0mL

Solution E 50.0mL

Solution F 10.0mL

Solution G 10.0mL

Solution B 1.0mL

Solution C 1.0mL Solution D 1.0mL

pH 7.3 ± 0.2 at 25°C

Solution A:

NH4Cl 0.25g

Solution B:

Solution C:

D(+)-Biotin 20.0mg

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

Preparation of Solution D: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Sparge with 100% N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Preparation of Solution E: Add NaHCO3 to distilled/deionized water and bring volume to 100.0mL Mix thoroughly Sparge with

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100% N2 gas mixture Autoclave for 15 min at 15 psi pressure–121°C.

Cool to 25°C

Solution F:

Na2-fumarate 3.2g

Na-acetate 0.8g

to 25°C

Solution G :

Na2S·9H2O 0.125g

Use: For the cultivation of Geovibrio thiophilus DSM 11263

Solution A 940.0mL

Solution E 50.0mL

Solution F 10.0mL

Solution G 10.0mL

Solution B 1.0mL

Solution C 1.0mL

Solution D 1.0mL

pH 7.3 ± 0.2 at 25°C

Solution A:

NaCl 1.0g

KCl 0.5g

MgCl2·6H2O 0.4g

NH4Cl 0.25g

KH2PO4 0.2g

CaCl2·2H2O 0.15g

Resazurin 0.5mg

25°C

Solution B:

FeCl2·4H2O 1.5g

CoCl2·6H2O 190.0mg

MnCl2·4H2O 100.0mg

ZnCl2 70.0mg

NiCl2·6H2O 24.0mg

Solution C:

D(+)-Biotin 20.0mg

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Solution F:

Na-glycolate 2.0g Yeast extract 1.0g

Preparation of Solution F: Add components to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100%

to 25°C

Na2S·9H2O 0.125g

Use: For the cultivation of Clostridium sp DSM 11261

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Solution A 940.0mL

Solution E 50.0mL

Solution F 10.0mL

Solution G 10.0mL

Solution B 1.0mL

Solution C 1.0mL

Solution D 1.0mL

pH 7.3 ± 0.2 at 25°C

Solution A:

NaCl 1.0g

KCl 0.5g

MgCl2·6H2O 0.4g

NH4Cl 0.25g

KH2PO4 0.2g

CaCl2·2H2O 0.15g

Resazurin 0.5mg

25°C

Solution B:

FeCl2·4H2O 1.5g

CoCl2·6H2O 190.0mg

MnCl2·4H2O 100.0mg

ZnCl2 70.0mg

NiCl2·6H2O 24.0mg

H3BO3 6.0mg

CuCl2·2H2O 2.0mg

Cool to 25°C

Solution C:

Vitamin B12 100.0mg

D(+)-Biotin 20.0mg

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

Preparation of Solution D: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Sparge with 100% N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Preparation of Solution E: Add NaHCO3 to distilled/deionized wa-ter and bring volume to 100.0mL Mix thoroughly Sparge with 100% N2 gas mixture Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C

Solution F:

Na-3-hydroxybutyrate 3.0g Yeast extract 1.0g

Preparation of Solution F: Add components to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100%

to 25°C

Na2S·9H2O 0.125g

Use: For the cultivation of unclassified bacterium DSM 11262

pH 7.3 ± 0.2 at 25°C

Solution A:

Na2SO4 2.8 g NaCl 1.0g KCl 0.5g MgCl2·6H2O 0.4g

NH4Cl 0.25g

Preparation of Solution A: Prepare under 80% N2 + 20% CO2 gas atmosphere Add components to distilled/deionized water and bring vol-ume to 940.0mL Mix thoroughly Adjust pH to 7.2 Sparge with 80% N2

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25°C

Solution B:

FeCl2·4H2O 1.5g

CoCl2·6H2O 190.0mg

MnCl2·4H2O 100.0mg

ZnCl2 70.0mg

NiCl2·6H2O 24.0mg

H3BO3 6.0mg

CuCl2·2H2O 2.0mg

Cool to 25°C

Solution C:

Vitamin B12 100.0mg

D(+)-Biotin 20.0mg

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Cool to 25°C

Solution F:

Syringic acid 0.6 g

Preparation of Solution F: Add syringic acid to distilled/deionized

water and bring volume to 10.0mL Mix thoroughly Adjust pH to 7.0

Sparge with 100% N2 gas mixture Autoclave for 15 min at 15 psi

pres-sure–121°C Cool to 25°C

Solution G :

Na2S·9H2O 0.25g

pH should be 7.2–7.4 Prior to inoculation the completed medium should equilibrate overnight

Use: For the cultivation of Parasporobacterium paucivorans DSM

15970

Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution H 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL

pH 6.5–7.0 at 25°C

Solution A:

NH4Cl 0.25g

Solution B:

Solution C:

Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H2O 200.0mg

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Vitamin B12 100.0mg

D(+)-Biotin 20.0mg

Solution D:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Cool to 25°C

Solution F:

Trimethylamine hydrochloride 2.0g

Methanol 0.6g

to 25°C

Solution G :

Na2S·9H2O 0.125g

Preparation of Solution G : Add Na2S·9H2O to distilled/deionized

Solution H:

FeSO4·6H2O 0.02g

Preparation of Solution H: Add FeSO4·6H2O to

distilled/deion-ized water and bring volume to 10.0mL Mix thoroughly Sparge with

Cool to 25°C

pH should be 6.5–7.0 After inoculation aseptically inject 0.1mL of

so-lution H for each 10.0mL of medium

Use: For the cultivation of Methanomethylovorans hollandica DSM

15978

Modified Gorodkowa Agar

Agar 20.0g Peptic digest of animal tissue 10.0g NaCl 5.0g Glucose 1.0g

pH 6.8 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring

to boiling with frequent agitation Distribute into tubes or flasks Auto-clave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes

or leave in tubes

Use: For promoting sporulation of yeasts

Modified ISP5 Medium (DSMZ Medium 993b)

Composition per liter:

KCl 100.0 Agar 20.0g Glycerol 10.0g Yeast extract 5.0g

L-asparagine, anhydrous 1.0g

K2HPO4, anhydrous 1.0g Trace elements solution 1.0mL

pH 7.2 ± 0.2 at 25°C

Trace Elements Solution:

FeSO4·7H2O 0.1g MnCl2·4H2O 0.1g ZnSO4·7H2O 0.1g

Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 100.0mL Mix

thorough-ly

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2 Distribute into tubes or flasks Gently heat while stirring and bring to boiling Mix thoroughly Autoclave for 15 min at 15 psi pressure– 121°C Pour into Petri dishes or leave in tubes

Use: For the cultivation of Nesterenkonia xinjiangensis and

Micro-bacterium halotolerans.

Modified ISP5 Medium with Sodium Chloride

(DSMZ Medium 993c)

Composition per liter:

NaCl 100.0g Agar 20.0g Glycerol 10.0g

L-asparagine, anhydrous 1.0g

K2HPO4, anhydrous 1.0g Trace elements solution 1.0mL

pH 7.2 ± 0.2 at 25°C

Trace Elements Solution:

FeSO4·7H2O 0.1g MnCl2·4H2O 0.1g ZnSO4·7H2O 0.1g

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