Team:UNITN-Trento/Notebook/Labposts/07/06

From 2013.igem.org

{"date" : "2013-07-02","author" : "fabio-viola","title" : "BACILLUS SUBTILIS COMES BACK TO LIFE!","content" : " in order to propagate B. subtilis from the pellet that we purchased (strain ind- tyr+ B.subtilis ATCC 23857) , first we needed to obtain the perfect medium for its growth. We prepared Nutrient Broth for liquid cultures and Nutrient Broth + Agar for plates, following ATCC PROTOCOLS. So we decided to have them in both a STARCH-added version and a version without starch.
Nutrient agar (100 ml) : 0,8 g nutrient broth: 1,5g Agar; water to 100 ml.
Nutrient agar + starch (100 ml) : 0,8 g nutrient broth: 1,5g Agar; 2ml starch from potato, water up to 100 ml.
Nutrient broth (250 ml): 2 g nutrient broth; 250 ml water.
Nutrient broth + starch (250 ml): 2 g nutrient broth; 5 ml starch; water up to 250 ml;
To propagate di original bacillus pellet, we resuspended it in 1 ml of nutrient broth+starch and put this milliliter in 5 ml of nutrient broth+starch for a final 6 ml mother liquid culture. Then we made several other liquid cultures with different bacteria concentrations from the Mother (for each concentration we made both starch and non-starch liquid cultures). We put all in a shaker at 26 degrees. We plated also the same concentrations in several plates with both Nutrient agar + starch and Nutrient agar alone, and put them at 26. Just out of curiosity we decided to put some plates and some liquid cultures at 37 degrees.","tags" : " Bacillus subtilis "}