SPP

SPP is a richer nutrient medium than NEFF and cells tend to exhibit faster doubling times.

1XSPP can be made by diluting 10XSPP stock medium or by making it fresh as outlined below.

Working Concentration: Proteose Peptone 2%, Yeast Extract  0.1%, Glucose 0.2%, FeCl3 33 uM

Method

 * 1) Fill beaker with 800 mL of ddH2O.
 * 2) Add all the dry ingredients and stir to mix well.
 * 3) Bring up the volume to 1 L.
 * 4) Add 0.2 g FeCl3-6H2O to the flask.
 * 5) Pour stock medium into spin bottles (250 mL capacity) and centrifuge at 5,000g for 20 minutes.
 * 6) Follow up by filtering through a Whatman filter apper. Immediately decant the medium into clean bottles and freeze. Do not sterilize the medium by autoclaving as the high amount of proteose peptone and glucose will get burnt during the autoclave cycle leaving a black residue.

Method

 * 1) Start again with 3/4 volume of the flask filled with ddH2O.
 * 2) Measure out the fry inredients and them to the beaker. Mix well by stirring.
 * 3) Add ddH2O to final volume of 1 L.
 * 4) Add 0.02 g of FeCl3-6H2O to the flask.
 * 5) Sterilize the medium by autoclaving.