Team:Purdue/MS media preparation

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Latest revision as of 19:37, 8 July 2010

Materials

For one liter of 0.5 [http://en.wikipedia.org/wiki/Murashige_and_Skoog_medium MS]:

2.165 g MS salt
15 g sucrose (1.5%)
12 g agar (1.2%)
1 N NaOH or KOH

Procedure

Use container that is twice as large as solution volume (ex. for 1 L of media, use a 2 L flask). Add MS salt and sucrose to 900mL distilled water. Adjust pH to 5.7 using 1 N NaOH or KOH solutions. Finally, add agar (it is important to add agar after balancing pH, as to not coat the pH meter with agar). There is no need to dissolve the agar prior to autoclaving. Cover flask/container with double layer aluminum foil. Place in autoclave and select P6-Liquid-Long Cycle. Total time for autoclave is 60 min. Leave aluminum foil on and keep the agar in a 55 degree bath until ready for use.

Courtesy of Mike Poling, Lab Technician: Horticulture & Landscape Architecture, Purdue University.

@@ Line 1: / Line 1: @@
-[[Arabidopsis grown on 0.5 MS media]]
+==Materials==
-For one liter of 0.5 MS:
+For one liter of 0.5 [http://en.wikipedia.org/wiki/Murashige_and_Skoog_medium MS]:
-.165 gm MS salt + 15 gm sucrose (1.5%) + 12 gm agar (1.2%). Adjust the pH to 5.7 with I N NaOH.
+*2.165 g MS salt
+*15 g sucrose (1.5%)
+*12 g agar (1.2%)
+*1 N NaOH or KOH
-[[Procedure]]
+==Procedure==
-Add MS salt and sucrose to 900mL distilled water. Adjust pH to 5.7. LASTLY, add agar.
+Use container that is twice as large as solution volume (ex. for 1 L of media, use a 2 L flask). Add MS salt and sucrose to 900mL distilled water. Adjust pH to 5.7 using 1 N NaOH or KOH solutions.  Finally, add agar (it is important to add agar after balancing pH, as to not coat the pH meter with agar).  There is no need to dissolve the agar prior to autoclaving.  Cover flask/container with double layer aluminum foil. Place in autoclave and select P6-Liquid-Long Cycle. Total time for autoclave is 60 min. Leave aluminum foil on and keep the agar in a 55 degree bath until ready for use.
-[[AutoClave Notes:]]
+Courtesy of Mike Poling, Lab Technician: Horticulture & Landscape Architecture, Purdue University.
-Use container that is twice as large as solution volume (ex. for 500 mL solution one should use 1000mL flask). Cover flask/container with double layer aluminum foil. Place in AutoClave and select P6-Liquid-Long Cycle. Total time for AutoClave is 60 min.