{"medium":{"id":346,"name":"DESULFALLAS MEDIUM","complex_medium":"yes","min_pH":7.2,"max_pH":7.5,"source":"DSMZ","link":"https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium346.pdf"},"solutions":[{"id":662,"name":"Main sol. 346","volume":1012,"recipe":[{"recipe No.":1,"solution":"Solution A","solution_id":663,"amount":966,"unit":"ml","optional":"no"},{"recipe No.":2,"solution":"Solution B","solution_id":664,"amount":20,"unit":"ml","optional":"no"},{"recipe No.":3,"solution":"Solution C","solution_id":665,"amount":11,"unit":"ml","optional":"no"},{"recipe No.":4,"solution":"Solution D","solution_id":666,"amount":10,"unit":"ml","optional":"no"},{"recipe No.":5,"solution":"Solution E","solution_id":667,"amount":1,"unit":"ml","optional":"no"},{"recipe No.":6,"solution":"Solution F","solution_id":668,"amount":1,"unit":"ml","optional":"no"},{"recipe No.":7,"solution":"Solution G","solution_id":669,"amount":1,"unit":"ml","optional":"no"},{"recipe No.":8,"solution":"Solution H","solution_id":670,"amount":1.7,"unit":"ml","optional":"no"}],"steps":[{"step":"Sparge solution A with 100% N2 gas for 30 - 45 min to make it anoxic. Thereafter, dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Anoxic stock solutions C, D, E, F, and H are prepared under 100% N2 gas atmosphere and solution G under 80% N2 and 20% CO2 gas atmosphere. Filter sterilize solutions D, F and G. To complete the medium appropriate amounts of solutions B to H are added to the sterile solution A in the sequence as indicated. Adjust pH of the complete medium to 7.2 - 7.5, if necessary."}],"equipment":{"4840":"Gassing station","4841":"Hungate tubes","4842":"Autoclave","4843":"Filter","5018":"Centrifuge"},"main solution":"yes"},{"id":663,"name":"Solution A","volume":966,"recipe":[{"recipe No.":1,"compound":"Na2SO4","compound_id":110,"amount":3,"unit":"g","g_l":3.10559,"mmol_l":21.8642,"optional":"no"},{"recipe No.":2,"compound":"KH2PO4","compound_id":11,"amount":0.2,"unit":"g","g_l":0.207039,"mmol_l":1.5214,"optional":"no"},{"recipe No.":3,"compound":"NH4Cl","compound_id":42,"amount":0.3,"unit":"g","g_l":0.310559,"mmol_l":5.80593,"optional":"no"},{"recipe No.":4,"compound":"CaCl2 x 2 H2O","compound_id":7,"amount":0.15,"unit":"g","g_l":0.15528,"mmol_l":1.05622,"optional":"no"},{"recipe No.":5,"compound":"MgCl2 x 6 H2O","compound_id":79,"amount":0.4,"unit":"g","g_l":0.414079,"mmol_l":2.03676,"optional":"no"},{"recipe No.":6,"compound":"NaCl","compound_id":43,"amount":1,"unit":"g","g_l":1.0352,"mmol_l":17.7138,"optional":"no"},{"recipe No.":7,"solution":"Trace element solution SL-10","solution_id":595,"amount":1,"unit":"ml","optional":"no"},{"recipe No.":8,"compound":"Sodium resazurin","compound_id":1852,"attribute":"0.1% w/v","amount":0.5,"unit":"ml","g_l":0.000517598,"mmol_l":0.00206075,"optional":"no"},{"recipe No.":9,"compound":"Distilled water","compound_id":4,"amount":965,"unit":"ml","optional":"no"}],"main solution":"no"},{"id":664,"name":"Solution B","volume":20,"recipe":[{"recipe No.":1,"compound":"NaHCO3","compound_id":55,"amount":1,"unit":"g","g_l":50,"mmol_l":595.188,"optional":"no"},{"recipe No.":2,"compound":"Distilled water","compound_id":4,"amount":20,"unit":"ml","optional":"no"}],"main solution":"no"},{"id":665,"name":"Solution C","volume":11,"recipe":[{"recipe No.":1,"compound":"Ethanol","compound_id":201,"amount":1,"unit":"ml","optional":"no"},{"recipe No.":2,"compound":"Distilled water","compound_id":4,"amount":10,"unit":"ml","optional":"no"}],"main solution":"no"},{"id":666,"name":"Solution D","volume":10,"recipe":[{"recipe No.":1,"compound":"Na-benzoate","compound_id":295,"amount":0.7,"unit":"g","g_l":70,"mmol_l":485.764,"optional":"no"},{"recipe No.":2,"compound":"Distilled water","compound_id":4,"amount":10,"unit":"ml","optional":"no"}],"main solution":"no"},{"id":667,"name":"Solution E","volume":1,"recipe":[{"recipe No.":1,"solution":"Clarified rumen fluid","solution_id":2618,"condition":"","amount":1,"unit":"ml","optional":"no"}],"main solution":"no"},{"id":668,"name":"Solution F","volume":1,"recipe":[{"recipe No.":2,"solution":"Wolin's vitamin solution (10x)","solution_id":5980,"amount":1,"unit":"ml","optional":"no"}],"main solution":"no"},{"id":669,"name":"Solution G","volume":1,"recipe":[{"recipe No.":3,"solution":"Na-dithionite solution (5% w/v)","solution_id":6262,"amount":1,"unit":"ml","optional":"no"}],"main solution":"no"},{"id":670,"name":"Solution H","volume":2,"recipe":[{"recipe No.":1,"compound":"Na2S x 9 H2O","compound_id":54,"amount":50,"unit":"mg","g_l":25,"mmol_l":104.088,"optional":"no"},{"recipe No.":2,"compound":"Distilled water","compound_id":4,"amount":1.7,"unit":"ml","optional":"no"}],"main solution":"no"},{"id":595,"name":"Trace element solution SL-10","volume":1000,"recipe":[{"recipe No.":1,"compound":"HCl","compound_id":68,"attribute":"25%","amount":10,"unit":"ml","g_l":2.5,"mmol_l":68.5683,"optional":"no"},{"recipe No.":2,"compound":"FeCl2 x 4 H2O","compound_id":260,"amount":1.5,"unit":"g","g_l":1.5,"mmol_l":7.54482,"optional":"no"},{"recipe No.":3,"compound":"ZnCl2","compound_id":64,"amount":70,"unit":"mg","g_l":0.07,"mmol_l":0.513573,"optional":"no"},{"recipe No.":4,"compound":"MnCl2 x 4 H2O","compound_id":36,"amount":100,"unit":"mg","g_l":0.1,"mmol_l":0.505293,"optional":"no"},{"recipe No.":5,"compound":"H3BO3","compound_id":37,"amount":6,"unit":"mg","g_l":0.006,"mmol_l":0.0970246,"optional":"no"},{"recipe No.":6,"compound":"CoCl2 x 6 H2O","compound_id":38,"amount":190,"unit":"mg","g_l":0.19,"mmol_l":1.46335,"optional":"no"},{"recipe No.":7,"compound":"CuCl2 x 2 H2O","compound_id":39,"amount":2,"unit":"mg","g_l":0.002,"mmol_l":0.0117314,"optional":"no"},{"recipe No.":8,"compound":"NiCl2 x 6 H2O","compound_id":40,"amount":24,"unit":"mg","g_l":0.024,"mmol_l":0.185187,"optional":"no"},{"recipe No.":9,"compound":"Na2MoO4 x 2 H2O","compound_id":9,"amount":36,"unit":"mg","g_l":0.036,"mmol_l":0.148791,"optional":"no"},{"recipe No.":10,"compound":"Distilled water","compound_id":4,"amount":990,"unit":"ml","optional":"no"}],"steps":[{"step":"First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml."}],"Sol. No.":2,"main solution":"no"},{"id":2618,"name":"Clarified rumen fluid","volume":1000,"steps":[{"step":"Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121\u00b0C 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\u00b0C."}],"Sol. No.":6,"main solution":"no"},{"id":5980,"name":"Wolin's vitamin solution (10x)","volume":1000,"recipe":[{"recipe No.":1,"compound":"Biotin","compound_id":46,"amount":20,"unit":"mg","g_l":0.02,"mmol_l":0.0818632,"optional":"no"},{"recipe No.":2,"compound":"Folic acid","compound_id":138,"amount":20,"unit":"mg","g_l":0.02,"mmol_l":0.0453104,"optional":"no"},{"recipe No.":3,"compound":"Pyridoxine hydrochloride","compound_id":519,"amount":100,"unit":"mg","g_l":0.1,"mmol_l":0.486287,"optional":"no"},{"recipe No.":4,"compound":"Thiamine HCl","compound_id":1207,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.148236,"optional":"no"},{"recipe No.":5,"compound":"Riboflavin","compound_id":136,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.132837,"optional":"no"},{"recipe No.":6,"compound":"Nicotinic acid","compound_id":139,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.406141,"optional":"no"},{"recipe No.":7,"compound":"Calcium D-(+)-pantothenate","compound_id":1512,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.209846,"optional":"no"},{"recipe No.":8,"compound":"Vitamin B12","compound_id":18,"amount":1,"unit":"mg","g_l":0.001,"mmol_l":0.00073779,"optional":"no"},{"recipe No.":9,"compound":"p-Aminobenzoic acid","compound_id":47,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.364591,"optional":"no"},{"recipe No.":10,"compound":"(DL)-alpha-Lipoic acid","compound_id":252,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.24233,"optional":"no"},{"recipe No.":11,"compound":"Distilled water","compound_id":4,"amount":1000,"unit":"ml","optional":"no"}],"main solution":"no"},{"id":6262,"name":"Na-dithionite solution (5% w/v)","volume":10,"recipe":[{"recipe No.":1,"compound":"NaOH","compound_id":256,"attribute":"1 N","amount":0.5,"unit":"ml","optional":"no"},{"recipe No.":2,"compound":"Na2S2O4","compound_id":653,"amount":0.5,"unit":"g","g_l":50,"mmol_l":287.175,"optional":"no"},{"recipe No.":3,"compound":"Distilled water","compound_id":4,"amount":9.5,"unit":"ml","optional":"no"}],"steps":[{"step":"Add 1 N NaOH to distilled water and make the solution anoxic by sparging with 100% N2 gas. Then dissolve the Na-dithionite and filter-sterilize the solution into anoxic Hungate tubes. The final pH of the solution should be between 9 and 10. Store the prepared solution in a dark and cool place. Prepare only small amounts of stock solution, as Na-dithionite decomposes rapidly."}],"main solution":"no"}]}