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Team:Purdue/MS media preparation
From 2010.igem.org
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For one liter of 0.5 [http://en.wikipedia.org/wiki/Murashige_and_Skoog_medium MS]: | For one liter of 0.5 [http://en.wikipedia.org/wiki/Murashige_and_Skoog_medium MS]: | ||
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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. | 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. | ||
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| + | Courtesy of Mike Poling, Lab Technician: Horticulture & Landscape Architecture, Purdue University. | ||
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.
