<Features>
 
JM Medium is optimized for gut microbial culture which universially supports the growth of gut microbes.
 
By using JM medium, we were able to construct a gut microbe library consistin of more than 2,200 species.
 
Research has proven that JM media has dramatically stimulated gut microbe's growth.
 
An overnight culture on JM agar medium can take sufficient for colony counting.
 
Through JM medium, You can also discover newly gut microbes.
 
 
 
<Constitution and Function>
 
Base powder : Composed of factors that support intestinal microbial culture.
 
JM supplement : Mixture consisting of 19 plants to the basal medium dramatically stimulated the growth of gut microbes. 
(* JM Supplement is an "insoluble" component, so it is normal not to dissolve in water.)
 
 
SCFA solution : Supplements for gut microbial culture. *Purchase separately
 
Vitamin solution : Supplements for gut microbial culture. *Purchase separately
 
Trace elements solution : Supplements for gut microbial culture. *Purchase separately
 
Hemin solution : Supplements for gut microbial culture. *Purchase separately
 
 
<Directions>
 
Suspend 31g of the base powder in 900 mL of purified water.
Add Glycerol 2 mL, Hemin solution 0.2 mL, Trance elements solution 1 mL and SCFA solution 0.578 mL.
Add 31g of the JM vegetable powder.
Mix thoroughly the ingredients with magnetic stirrer.
Adjust the pH to 7.4 with 10N NaOH.
For agar medium, add 15g of agar.
Adjust the final volume of the solution to 1 L with water.
Warm slighly to completely dissove the powder.
Sterilize by autoclave for 20 min at 15 psi (1.05 kg/cm2) on liquid cycle.
Coll the medium below 40 ℃.
Add 50 mL pig blood and 2 mL of the vitamin solution.
For agar medium, pour the sterile medium in each petri dish and leave in the hood for 30 minutes to cool and solidify.
Use the broth or agar medium after removing oxygen by incubating the medium in an anaerobic incbator for overnight.
 
 
<Preparation of stock solution>
 
1. Hemin stock solution
Dissolve 50 mg Hemin in 1 mL 1 N NaOH.
Make up to 100 mL with purified water.
Sterilize the solution with a 0.22 µm filter.
 
2. Trace element stock solution
Weigh the following ingredients and mix them together in 900 mL of purified water, allowing them to dissove completely.
Sterilize the solution with a 0.22 µm filter.
MnCl2 x 4 H2O-------------------0.72 g
FeSO4 x 7 H2O--------------------0.2 g
CoCl2 x 6 H2O---------------------0.2 g
ZnSO4 x 7H2O--------------------0.2 g
Na2MoO4 x 2 H2O-------------0.02 g
NiCl2----------------------------------0.1 g
CuSO4 x 5 H2O-----------------0.02 g
H3BO3-----------------------------0.02 g
MgSO4. 7H2O--------------------0.6 g
CaCl2. 2H2O-----------------------0.6 g
(NH4)2SO4---------------------------1 g
 
3. SCFA (short chain fatty acid) stock solution
Mix the following ingredients in air hood and store at room temperature.
Sterilize the solution with a 0.22 µm filter.
Acetic acid---------------------------15 mL
Propionic acid------------------------6 mL
n-Valeric acid--------------------------1 mL
Iso-valeric acid------------------------1 mL
Iso-butyric acid-----------------------1 mL
 
4. Vitamin stock solution
Weight the following ingredients and mix them together in 900 mL of purified water, allowing them to dissolve completely.
Sterilize the solution with a 0.22 µm filter.
Biotin----------------------------------0.02 g
Cyanocobalamin------------------0.02 g
P-amino benzoic acid------------0.03 g
Folic acid-----------------------------0.05 g
Pyridoxine.HCl----------------------0.15 g
Thiamin. HCL------------------------0.05 g
Riboflavin-----------------------------0.05 g
L-ascorbic acid----------------------0.05 g
Coenzyme Q-------------------------0.04 g
L-glutamine---------------------------0.02 g
Trimethylglycine--------------------0.03 g
Inositol---------------------------------0.03 g
Niacin-----------------------------------0.03 g
L-carnitine-----------------------------0.03 g
Methylsulfonylmethane----------0.03 g
Vitamin K1----------------------------0.03 g
 
 
 
 
 
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