Editing Nutrient Broth (section)

== Culture of&nbsp;Bacillus subtilis  ==

=== ATCC Protocol ===

ATCC® Number: 6633D-5 ™<br>Deposited As Bacillus subtilis (Ehrenberg) Cohn<br>Strain Designations: Genomic DNA from strain NRS 231 [ATCC 6633] 

ATCC Medium: 415<br>Potato Extract Agar/Broth<br>For Solid Media:<br>Nutrient Agar .......................................23.0 g<br>Potato Extract (see below)......................20.0 ml<br>DI Water..............................................980.0 mL<br>Autoclave at 121 ºC<br>To prepare potato extract, boil 300 g peeled, diced potatoes in 500 ml distilled water for&nbsp;half an hour. Decant the infusion and use in the above medium. (Put through cheese&nbsp;cloth a few times.)<br>For Broth<br>Nutrient Broth....................................8.0 g<br>Potato Extract (see above) .................20.0 mL<br>DI Water.........................................980.0 mL<br>Autoclave at 121 ºC 

Growth Conditions <br>Temperature: 26°C<br>Atmosphere: Aerobic 



'''http://iwi.eldoc.ub.rug.nl/FILES/root/2011/PLoSOneBlom/2011PLoSOneBlom.pdf'''

TY medium (10 g/L trypton, 5 g/L yeast extract, 5 g/L NaCl) in batch mode&nbsp;at 37uC in a 16 L fermentor using a working volume of 12 L.<br>Cultures were aerated at 40 L/min and 400 rpm



=== Web Protocols ===

'''1. '''Bacillus subtilis 168 wild type species, and some other bacillus species. Growth conditions are simple, first try to grow on 0.5 LB agar plate ( 1% tryptone, 0.5% Yeast extract, 0.5% NaCl and Agar 1.5% ), after getting growth on LB agar plate, (for safer side just check morphology of Bacillus on plate). Inoculate single colony in 0.5 LB medium (no agar). 

'''2.'''&nbsp;&nbsp;B-Medium: 8 g Nutrient Broth (Difco) per l, autoclaved.

Add per 100 ml NB:<br>1. 0.5 ml Metal Mix (14 ml 1 M Ca-chloride; 10 ml 0.1 M Mn chloride; 20 ml 1 M Mg chloride; 56 ml water), sterile-filtered or made from pre-sterilized stock solutions<br>2. 0.05 ml Fe-solution (32.4 mg Fe(III)-chloride; 1 ml 1 M HCl in 100 ml water), sterile-filtered<br>3. 5 ml Phosphate buffer (95 g K2HPO4 x 3 H2O, 124 g KH2PO4, ad 1l water), autoclaved

'''3. &nbsp;'''1% Dextrose, 0.5% peptone and 0.45% yeast extract for the growth. pH should be 7-7.2 and temperature 30-37 degree Celcius. Huge growth will come within 12 hours.

'''4. &nbsp;'''&nbsp;Dextrose, 0.5% peptone and 0.45% yeast extract for the growth. pH should be 7-7.2 and temperature 30-37 degree Celcius. Huge growth will come within 12 hours.

[[Category:Buffer_Recipe]]