Medium data

Search for medium no. = [537].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

537   TREPONEMA ISOPTEROCOLA MEDIUM

NaCl  1.0     g
KCl  0.5     g
MgCl2·6H2O  0.4     g
CaCl2·2H2O  0.1     g
NH4Cl  0.3     g
KH2PO4  0.2     g
Na2SO4  0.15    g
Yeast extract  4.0     g
Peptone from casein  2.0     g
Resazurin  0.5     mg
Distilled water  1.0     L
Adjust pH to 7.2 - 7.4.
Prepare the medium anaerobically, dispense its 5 ml each in anaerobic tubes under a N2-CO2 atmosphere (80:20) and autoclave. Add the following solutions via injection from sterile anaerobic stock solutions per 5 ml:
Trace element solution SL-10* (see Medium No. 433), 10-fold diluted  0.05    ml
Vitamin solution** (see below), 10-fold diluted  0.05    ml
Co-factors solution*** (see below)  0.1     ml
NaHCO3 solution (10%, w/v)  0.3     ml
Sugar solution (250 mM each of xylose, maltose and cellobiose)  0.1     ml
Dithiothreitol (0.25 M)  0.05    ml
Vitamin solution:
Vitamin B12  100.0     mg
p-Aminobenzoic acid  80.0     mg
D(+)-Biotin  20.0     mg
Nicotinic acid  200.0     mg
Calcium pantothenate  100.0     mg
Pyridoxine·HCl  300.0     mg
Thiamine·HCl·2H2O  200.0     mg
Distilled water  1.0     L
Co-factors solution:
Pyridoxal·HCl  250.0     mg
Pyridoxal phosphate  250.0     mg
Calcium folinic acid  50.0     mg
β-NAD  50.0     mg
Coenzyme A  50.0     mg
FAD  50.0     mg
Nicotinamide  25.0     mg
Folic acid  2.5     mg
Riboflavin  0.5     mg
Hemin (dissolved separately in 10 mM NaOH)  65.0     mg
Thiamine pyrophosphate  2.5     g
Distilled water  1.0     L
Adjust pH to 7.0 with 1 N NaOH and filter-sterilize.



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