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.