Lidstrom:M9: Difference between revisions

Revision as of 10:13, 11 September 2012

Make sure autoclaved water for diluting the M9, etc. is warm. M9 salts bottle suggests 40-50 deg C. I keep a bottle of autoclaved water for preparing this media in the 65 deg C incubator, so it's ready when I need it.

Add 200mL 5X M9 salts solution to ~800mL of distilled water
Add 15g of agar
Autoclave
After autoclaving, swirl to mix evenly and "temper" at room temperature until you can place your hand on the flask for 2 second
Add 2mL of 1M MgSO4, 0.1mL of 1M CaCl2 + carbon source of choice

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-Make sure autoclaved water for diluting the M9, etc. is warm. M9 salts bottle suggests 40-50 deg C. I keep a bottle of autoclaved water for preparing this media in the 65 deg C incubator, so it's ready when I need it.
+==5x M9 Salts==
+*Using premixed Difco 5x right now
+==M9 (for 1 L)==
+# Add 100 uL 1M CaCl2 to 750 mL autoclaved, warm water (see note)
+# Add 2 mL 1M MgSo4
+# Add 200 mL 5x M9 salts
+# Add carbon source
+# Add any other supplements
+# Add autoclaved water to 1L
+*Make sure autoclaved water for diluting the M9, etc. is warm. M9 salts bottle suggests 40-50 deg C. I keep a bottle of autoclaved water for preparing this media in the 65 deg C incubator, so it's ready when I need it.
+==M9 Agar, 1L==
+# Add 200mL 5X M9 salts solution to ~800mL of distilled water
+# Add 15g of agar
+# Autoclave
+# After autoclaving, swirl to mix evenly and "temper" at room temperature until you can place your hand on the flask for 2 second
+# Add 2mL of 1M MgSO4, 0.1mL of 1M CaCl2 + carbon source of choice
+*[http://www.madsci.org/~lynn/micro/techniques/pour_plates/MINagar.html Mad Sci Protocol]
 *[http://www.thelabrat.com/protocols/m9minimal.shtml Lab Rat Protocol]