Editing Nutrient Broth

== Broth  ==

Mix 1 volume of fresh ground beef with 1 volume of tapwater and let stand overnight in refrigerator or room temperature. Heat to boiling, with stirring, to coagulate proteins. Filter through cloth. Discard the meat. If you worked cleanly and used refrigerator temperature for the extraction, the discarded meat is safe for human food. Add 1% peptone and 0.5% table salt (sodium chloride). Adjust pH to neutral using dilute NaOH (1.0 Normal is suggested) Heat to boiling again and filter thru filter paper if perfectly clear Broth is wanted. Dispense into tubes, plug with cotton, and autoclave. 

== E Broth  ==

Eddleman Broth is simpler for your home micro lab and many bacteria will grow very well on it, but the scum, sediment, and other results of bacterial growth will differ somewhat from the results on classic broth. Some bacteria grow better if you omit the salt. Salt (5 grams per liter) is used added if we are growing bacteria from human gut or decaying meat, but the bacteria usually grow fine without the salt. We will omit peptone because you don't have it. Use ground beef, or ground liver, or any kind of meat you wish. 

Mix 1/2 pound of fresh ground meat with 1 liter of tapwater and let stand several hours in the refrigerator, stir occasionaly to aid extraction of soluble proteins. Heat to boiling, with stirring, until the meat is cooked to coagulate proteins. Filter through a white cloth which is thick enough to remove all particles. Discard the meat. If you worked cleanly (clean pans and clean cloth) and used refrigerator temperature for the extraction, the discarded meat is safe for human food. Chill the broth in refrigerator so that the fat forms a hard floating layer. Discard the fat. Adjust the volume to one liter by adding tapwater. Do not adjust pH because you do not have a pH meter or pH paper and most bacteria will grow fine. If you wish add a 1/8 inch crushed limestone pebble to some of the culture tubes to see if there is any difference in growth of bacteria, or use powdered limestone or calcium carbonate. Dispense into tubes, plug with cotton, and autoclave in pressure cooker at 15 pounds pressure for 15 minutes. Let tubes cool in pressure cooker. 

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== Difco™ Nutrient Broth  ==

<br>Approximate Formula* Per Liter<br>Beef Extract .................................................................3.0 g<br>Peptone ......................................................................5.0 g 

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== 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]]