NGM

Submitted by kdorfman on Fri, 04/01/2016 - 16:31

Per Liter of medium (~75 plates):

  • 975 mL Water
  • 3 g NaCl
  • 2.5 g Peptone (Fisher BP1420-500 $78.80)
  • 17 g Bactoagar

Autoclave with stir bar inside

Cool to 55C in a 55C water bath

Add per L (see recipes in stock solutions):

  • 1 mL cholesterol (5 mg/mL in 95% EtOH)
  • 1 mL CaCl2 (1 M, STERILE)
  • 1 mL MgSO4 (1 M, STERILE)
  • 25 mL K-phosphate buffer (1M, pH 6.0, STERILE1)

Swirl flask to mix

Dispense 10 mL into each 60mm dish.

Stack 10 high

Let stand for ~48 hours for condensation to evaporate

Pack in sterilized plastic boxes.

To make NGM agar plates
at mm plate diameter
pour mL per plate
start with: mL water
add: g peptone, stir till dissolved
addg NaCL, stir till dissolved
add: g agar, leave stir bar in
autoclave for: minutes
add: mL 5mg/mL cholesterol
add: mL sterile 1M CaCl2
add: mL sterile 5mg/mL 1M MgSO4
add: mL sterile K-phosphate buffer , pH 6

put bottles on stir plate near sterile hood until handle-able

  1. 3.3 mL K2HPO4 + 21.7 mL KH2PO4 ↩︎