Overview
- External ontology
- mediadive.medium:161 (source link unavailable) — METHANOFOLLIS MEDIUM
- Applications
-
- Microbial cultivation
- pH
- 6.5
- Notes
- Source: DSMZ | Link: https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium161.pdf Superseded historical provenance, not a current formulation claim: Commercial Product: LB Medium (Luria-Bertani, Miller formulation) Supplier: Multiple suppliers (Difco, Sigma, etc.) Catalog: Multiple (L3522 Sigma, 244520 BD) Source: https://www.laboratorynotes.com/preparation-of-luria-bertani-lb-miller-broth/ Accessed: 2026-03-28 Constituent composition researched from product specification. Original nominal final volume is1001ml, while listed liquid inputs total1001.5ml. Use1.001 L for the ten main mass rates, rounded to six significant digits, not measured precision or assumed additive volume. Preserve exact stock batch inputs without inventing final stock denominators. Preserve preparation stages: anoxic main preparation, separate clarified-fluid procedure, mineral-stock titration and fatty-acid-stock titration. Existing preparation steps remain source-scoped; storage filling fractions are not medium concentrations. Complete inspected source rows define this reconstruction. Archive false LB expansions, flattened quantities and placeholders in append-only history. Preserve earlier supported evidence exactly; unrelated chemical annotations are not recertified.
Composition
flowchart LR
CultureMech_001100["methanofollis_medium"]
ing0_K2HPO4["K2HPO4"]
CultureMech_001100 --> ing0_K2HPO4
CHEBI_131527["CHEBI:131527"]
ing0_K2HPO4 -.-> CHEBI_131527
ing1_KH2PO4["KH2PO4"]
CultureMech_001100 --> ing1_KH2PO4
CHEBI_63036["CHEBI:63036"]
ing1_KH2PO4 -.-> CHEBI_63036
ing2__NH4_2SO4["(NH4)2SO4"]
CultureMech_001100 --> ing2__NH4_2SO4
CHEBI_62946["CHEBI:62946"]
ing2__NH4_2SO4 -.-> CHEBI_62946
ing3_NaCl["NaCl"]
CultureMech_001100 --> ing3_NaCl
CHEBI_26710["CHEBI:26710"]
ing3_NaCl -.-> CHEBI_26710
ing4_MgSO4_x_7_H2O["MgSO4 x 7 H2O"]
CultureMech_001100 --> ing4_MgSO4_x_7_H2O
CHEBI_31795["CHEBI:31795"]
ing4_MgSO4_x_7_H2O -.-> CHEBI_31795
ing5_CaCl2_x_2_H2O__0_1__w_v_["CaCl2 x 2 H2O (0.1% w/v)"]
CultureMech_001100 --> ing5_CaCl2_x_2_H2O__0_1__w_v_
CHEBI_86158["CHEBI:86158"]
ing5_CaCl2_x_2_H2O__0_1__w_v_ -.-> CHEBI_86158
ing6_FeSO4_x_7_H2O__0_1__w_v_in_0_1_N_H2SO4_["FeSO4 x 7 H2O (0.1% w/v in 0.1 N H2SO4)"]
CultureMech_001100 --> ing6_FeSO4_x_7_H2O__0_1__w_v_in_0_1_N_H2SO4_
CHEBI_75836["CHEBI:75836"]
ing6_FeSO4_x_7_H2O__0_1__w_v_in_0_1_N_H2SO4_ -.-> CHEBI_75836
ing7_Yeast_extract__DIFCO_["Yeast extract (DIFCO)"]
CultureMech_001100 --> ing7_Yeast_extract__DIFCO_
ing8_Trypticase__BBL_["Trypticase (BBL)"]
CultureMech_001100 --> ing8_Trypticase__BBL_
ing9_Sodium_resazurin__0_1__w_v_["Sodium resazurin (0.1% w/v)"]
CultureMech_001100 --> ing9_Sodium_resazurin__0_1__w_v_
ing10_NaHCO3["NaHCO3"]
CultureMech_001100 --> ing10_NaHCO3
CHEBI_32139["CHEBI:32139"]
ing10_NaHCO3 -.-> CHEBI_32139
ing11_L_Cysteine_HCl_x_H2O["L-Cysteine HCl x H2O"]
CultureMech_001100 --> ing11_L_Cysteine_HCl_x_H2O
CHEBI_91248["CHEBI:91248"]
ing11_L_Cysteine_HCl_x_H2O -.-> CHEBI_91248
ing12_Na2S_x_9_H2O["Na2S x 9 H2O"]
CultureMech_001100 --> ing12_Na2S_x_9_H2O
CHEBI_76209["CHEBI:76209"]
ing12_Na2S_x_9_H2O -.-> CHEBI_76209
ing13_Distilled_water["Distilled water"]
CultureMech_001100 --> ing13_Distilled_water
Ingredients (14)
| # | Ingredient | Concentration | Ontology | Notes |
|---|---|---|---|---|
| 1 |
K2HPO4
HO4P.2K 174.174 g/mol
|
0.299700 G_PER_L |
CHEBI:131527
dipotassium hydrogen phosphate
|
Original main formulation input: 0.30 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 0.30 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml |
| 2 |
KH2PO4
H2O4P.K 136.084 g/mol
|
0.299700 G_PER_L |
CHEBI:63036
potassium dihydrogen phosphate
|
Original main formulation input: 0.30 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 0.30 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml |
| 3 |
(NH4)2SO4
2H4N.O4S 132.141 g/mol
|
0.299700 G_PER_L |
CHEBI:62946
ammonium sulfate
|
Original main formulation input: 0.30 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 0.30 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml |
| 4 |
NaCl
Cl.Na 58.443 g/mol
|
0.599401 G_PER_L |
CHEBI:26710
sodium chloride
|
Superseded flattening history, not a current amount: [Merged 2 duplicates: 0.599401, 1.0] Original main formulation input: 0.60 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 0.60 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml |
| 5 |
MgSO4 x 7 H2O
7H2O.Mg.O4S 246.473 g/mol
|
0.129870 G_PER_L |
CHEBI:31795
magnesium sulfate heptahydrate
|
Superseded flattening history, not a current amount: [Merged 2 duplicates: 0.12987, 3.0] Original main formulation input: 0.13 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 0.13 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml |
| 6 |
CaCl2 x 2 H2O (0.1% w/v)
Ca.2Cl.2H2O 147.014 g/mol
|
0.008 L |
CHEBI:86158
calcium chloride dihydrate
|
Superseded flattening history, not a current amount: [Merged 2 duplicates: 0.00799201, 0.1] Original main formulation input: 8.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
| 7 |
FeSO4 x 7 H2O (0.1% w/v in 0.1 N H2SO4)
Fe.7H2O.O4S 278.013 g/mol
|
0.002 L |
CHEBI:75836
iron(2+) sulfate heptahydrate
|
Superseded flattening history, not a current amount: [Merged 2 duplicates: 0.001998, 0.1] Original main formulation input: 2.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
| 8 | Yeast extract (DIFCO) | 0.999001 G_PER_L | Original main formulation input: 1.00 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 1.00 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml | |
| 9 | Trypticase (BBL) | 0.999001 G_PER_L |
MICRO:0000175 (source link unavailable)
Trypticase peptone
|
Original main formulation input: 1.00 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 1.00 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml |
| 10 | Sodium resazurin (0.1% w/v) | 0.0005 L | Original main formulation input: 0.50 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose | |
| 11 |
NaHCO3
CHO3.Na 84.006 g/mol
|
1.99800 G_PER_L |
CHEBI:32139
sodium hydrogencarbonate
|
Original main formulation input: 2.00 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 2.00 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml |
| 12 |
L-Cysteine HCl x H2O
C3H7NO2S.H2O.HCl 175.637 g/mol
|
0.499500 G_PER_L |
CHEBI:91248
L-cysteine hydrochloride hydrate
|
Original main formulation input: 0.50 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 0.50 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml |
| 13 |
Na2S x 9 H2O
9H2O.2Na.S 240.182 g/mol
|
0.499500 G_PER_L |
CHEBI:76209
sodium sulfide nonahydrate
|
Original main formulation input: 0.50 g per preparation. Use explicitly stated nominal final volume 1001 ml = 1.001 L, rounded to six significant digits; not measured precision. Listed liquid inputs sum to 1001.5 ml, not a replacement denominator Calculation: 0.50 g / 1.001 L. Additional exact excerpt (page1 header): Final volume: 1001 ml |
| 14 | Distilled water | 0.66 L | Original main formulation input: 660.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
Preparation
- Dissolve ingredients (except bicarbonate, vitamins, cysteine and sulfide) and sparge medium for 30 - 45 min with 80% H2 and 20% CO2 gas mixture to make it anoxic. Add and dissolve bicarbonate and adjust pH to 6.8, then distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas atmosphere. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of complete medium to 6.5. For incubation use sterile 80% H2 and 20% CO2 gas mixture at two atmospheres of pressure.
- Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 min and then centrifuged at 27,000 g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min, dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C.
- First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.
- Adjust pH to 7.5 with concentrated NaOH.
Solutions
Clarified rumen fluid
Dose: 0.30 L
Original main formulation input: 300.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose
Procedure-only clarified fluid; no ingredient amounts are stated for its preparation. The 30% vial-fill instruction concerns storage, not concentration. No standalone CultureMech identity is inferred. Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is
filtered through muslin, autoclaved at 121°C for 15 min and then centrifuged at 27,000 g
for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min,
dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then
stored frozen at -20°C.
Modified Wolin's mineral solution
Dose: 0.01 L
| Ingredient | Concentration | Notes |
|---|---|---|
| Nitrilotriacetic acid | Original mineral stock input: 1.50 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| MgSO4 x 7 H2O | Superseded flattening history, not a current amount: [Merged 2 duplicates: 0.12987, 3.0] Original mineral stock input: 3.00 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| MnSO4 x H2O | Original mineral stock input: 0.50 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| NaCl | Superseded flattening history, not a current amount: [Merged 2 duplicates: 0.599401, 1.0] Original mineral stock input: 1.00 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| FeSO4 x 7 H2O | Superseded flattening history, not a current amount: [Merged 2 duplicates: 0.001998, 0.1] Original mineral stock input: 0.10 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| CoSO4 x 7 H2O | Original mineral stock input: 0.18 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| CaCl2 x 2 H2O | Superseded flattening history, not a current amount: [Merged 2 duplicates: 0.00799201, 0.1] Original mineral stock input: 0.10 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| ZnSO4 x 7 H2O | Original mineral stock input: 0.18 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| CuSO4 x 5 H2O | Original mineral stock input: 0.01 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| AlK(SO4)2 x 12 H2O | Original mineral stock input: 0.02 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| H3BO3 | Original mineral stock input: 0.01 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Na2MoO4 x 2 H2O | Original mineral stock input: 0.01 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| NiCl2 x 6 H2O | Original mineral stock input: 0.03 g per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Na2SeO3 x 5 H2O | Original mineral stock input: 0.30 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Na2WO4 x 2 H2O | Original mineral stock input: 0.40 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Distilled water | 1.00 L | Original mineral stock input: 1000.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
| KOH | Source identifies this titrant and pH target but gives no fixed amount; do not infer zero or a typical concentration |
Original main formulation input: 10.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose
Fatty acid mixture
Dose: 0.02 L
| Ingredient | Concentration | Notes |
|---|---|---|
| Isobutyric acid | 0.023 L | Original fatty-acid stock input: 23.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
| DL-2-Methylbutyric acid | 0.027 L | Original fatty-acid stock input: 27.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
| Valeric acid | 0.027 L | Original fatty-acid stock input: 27.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
| Isovaleric acid | 0.027 L | Original fatty-acid stock input: 27.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
| Distilled water | 0.896 L | Original fatty-acid stock input: 896.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
| NaOH | Source identifies this titrant and pH target but gives no fixed amount; do not infer zero or a typical concentration |
Original main formulation input: 20.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose
Wolin's vitamin solution (10x)
Dose: 0.001 L
| Ingredient | Concentration | Notes |
|---|---|---|
| Biotin | Original vitamin stock input: 20.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Folic acid | Original vitamin stock input: 20.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Pyridoxine hydrochloride | Original vitamin stock input: 100.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Thiamine HCl | Original vitamin stock input: 50.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Riboflavin | Original vitamin stock input: 50.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Nicotinic acid | Original vitamin stock input: 50.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Calcium D-(+)-pantothenate | Original vitamin stock input: 50.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Vitamin B12 | Original vitamin stock input: 1.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| p-Aminobenzoic acid | Original vitamin stock input: 50.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| (DL)-alpha-Lipoic acid | Original vitamin stock input: 50.00 mg per preparation. Retain literal batch mass and stage in evidenced notes; source water addition alone does not establish a final stock denominator | |
| Distilled water | 1.00 L | Original vitamin stock input: 1000.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose |
Original main formulation input: 1.00 ml per preparation. Exact liquid input per preparation, not a percentage, mass, measured final volume or universal working dose
Target organisms
No target organisms are recorded.
Provenance
| Timestamp | Curator | Action | Notes |
|---|---|---|---|
| 2026-01-26T17:53:10.372381Z | mediadive-import | Imported from MediaDive | Source: DSMZ, ID: 161 |
| 2026-01-28T19:48:34.417173Z | chebi-enrichment | Added CHEBI ontology terms to ingredients | Enriched using MicrobeMediaParam and MediaDive chemical mappings |
| 2026-02-06T05:10:44.598268Z | invalid-chebi-removal | Removed 1 invalid CHEBI ID(s) | Removed CHEBI IDs with invalid format (7+ digits) from MicrobeMediaParam source data |
| 2026-03-13T03:09:28.715510Z | data-quality-cleanup-v1.0 | Data quality fixes | Merged 4 duplicate ingredient(s) |
| 2026-03-13T06:19:01.193330Z | culturemech-id-assigner-v1.0 | Assigned CultureMech ID | Assigned stable identifier: CultureMech:001100 |
| 2026-03-17T06:30:38.000000Z | mediaingredientmech-enrichment-v1.0 | Added MediaIngredientMech links | Linked ingredients to MediaIngredientMech identifiers |
| 2026-03-17T19:45:12.884090+00:00 | normalize_media_names_to_snake_case.py | NAME_NORMALIZED | Normalized name from "METHANOFOLLIS MEDIUM" to "methanofollis_medium" for consistent matching |
| 2026-06-05T07:41:31.456065+00:00 | mim-legacy-to-chebi-migration-v1.0 | Refreshed legacy MediaIngredientMech links to CHEBI keying | Replaced 33 legacy mediaingredientmech_term link(s) with mediaingredientmech_chebi_term (id-safe: ingredient's own CHEBI + label). MIM deprecated the MediaIngredientMech:NNNNNN id scheme. |
| 2026-06-10T07:47:06.948554+00:00 | primary-term-label-canonicalization-v1.0 | Canonicalized primary term label to OBO canonical (OAK) | 4 primary term(s); ingredient name resolved by OAK sqlite:obo:chebi. |
| 2026-09-22T00:03:26.686152+00:00 | repair_rejected_peptone_groundings.py | CORRECTED_REJECTED_PEPTONE_GROUNDING | CultureMech #484. /ingredients/8/term/id: Trypticase CHEBI:78018 -> MICRO:0000175 (replace_term). Reviewed identities: https://github.com/CultureBotAI/MediaIngredientMech/tree/24562493919391df4468a4285b7c997960cc8868. Publication resolver: MIM label index 9f09e4fb97fb0e6cbbd6f25baca40b36512adb88. MediaDive source IDs and concentrations preserved. |
| 2026-10-09T09:34:31.181526+00:00 | codex | RESTORED_METHANOFOLLIS_SOURCE_FORMULATIONS_WITH_EVIDENCE | Restore distinct medium161 source formulations, stock scopes, input units and unquantified titrants with exact evidence. Preserve prior supported entries and unknown quantities; correct false duplicate direction and stale paths. No growth association added. |
| 2026-10-09T16:36:08.086606+00:00 | codex | WITHDREW_UNSUPPORTED_RESAZURIN_STOCK_PRIMARY_IDENTITIES | ingredients[9].term: Withdrawn unsupported exact primary identity CHEBI:8806. The quoted recipe specifies a sodium-resazurin stock, whereas the local ChEBI snapshot identifies CHEBI:8806 as sodium-free Resazurin (formula C12H7NO4, charge 0; http://purl.obolibrary.org/obo/CHEBI_8806). No replacement chemical identifier or absence of a suitable ontology term is asserted. The source name, 0.0005 L preparation input and all other quantities are preserved. Locator=page 1, main formulation; accessed=2026-10-09T08:54:14.964059+00:00; text SHA256=8cc1d165274c4c0ce7a215cbe458c48d9880b183c7d62cbefa2d54e637d7625a; PDF SHA256=0f4ea6194efcb146abe36366dd72ce890643e9cfe39bc947a9df7281959478e0. The immutable MIM label-index snapshot (SHA256=a36cd4683feb89e2fc2a1721dc15008d1e10c12044e89a36c27b6621a8aaf262) still resolves this exact label to CHEBI:8806 even without the local term. That publication-time identity conflict remains unresolved and needs an upstream mapping review; the resolver and snapshot have not been altered. Live ChEBI lookup was unavailable through the web tool; ontology-form comparison uses the local snapshot. |