Ferment It Yourself

Vitamin A	Vitamin B2	Vitamin B12
Phytase	Riboswitch	Differentiation on E. coli
Differentiation on S. cerevisiae	Manufacturing	Idli and Micro-organisms

July

Received Propionibacterium freudenreichii subsp. shermanii CIRM BIA1 from the INRA Rennes CIRM collection.
Made the following media:
- MEA (waking lyophilized bacteria up), 1 L
  
  Product Quantity
  
  BHI broth 37 g
  
  Soja peptone 5 g
  
  Yeast extract 5 g
  
  Glucose 3 g
  
  Water Fill to 1 L
- YEL (propionibacteria growth medium), 1 L
  
  Product Quantity
  
  Sodium lactate (60% syrup) 21.4 g
  
  Tryptone 10 g
  
  Yeast extract 10 g
  
  K₂HPO₄, 3 H₂O 328 mg
  
  MnSO₄, H₂O 56 mg
  
  Water Fill to 1 L
Successfully grew P. freudenreichii in MEA at 30°C, in aerobic conditions.
Made a glycerol stock of it: g15.53.

Plated from the original P. freudenreichii glycerol stock on MEA. Did not grow.
For now, propioni grows only in MEA liquid (does not grow on MEA+agar nor YEL+agar

.

Made a gram coloration to assess whether the strain I have is gram positive, as P. freudenreichii.
They seemed purple (as Gram positive bacteria).
Need further checking.

Tried to extract the genomic DNA to sequence the 16s rRNA.
Problem: followed only the short sheet in the Qiagen DNeasy Blood & Tissue extraction kit, which was not made for gram-positive bacteria.
No DNA extracted at all.

Re-used the kit, this time adding lysozyme (10 mg/ml) to degrade the cell wall, but still no DNA out of the extraction.

Product
20 mM Tris-Cl, pH 8.0
2 mM sodium EDTA
1.2% Triton X-100
Immediately before use, add lysozyme to 20 mg/ml