Lidstrom:MM1 recipe: Difference between revisions
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#Prepare solution #2 mixture (in a 1L bottle) and add a stir bar to assure that all components are dissolved well. | #Prepare solution #2 mixture (in a 1L bottle) and add a stir bar to assure that all components are dissolved well. | ||
##'''Do not let this solution sit long before autoclaving.''' This solution should be light pink/orange by the time you combine it with solution #1. | ##'''Do not let this solution sit long before autoclaving.''' This solution should be light pink/orange by the time you combine it with solution #1. | ||
#Before autoclaving, | #Before autoclaving, combine bottles into a 2L bottle. | ||
#Autoclave as soon as possible or else things will precipitate! | |||
#Autoclave as soon as possible! | |||
== MM1 liquid recipe == | == MM1 liquid recipe == |
Revision as of 15:48, 5 March 2015
Back to Lidstrom:Protocols
Trace Elements Recipe & Tips
- Total volume: 1L (adjust accordingly)
- Solution #1 (500 mL ddH2O):
- 4.4 g ZnSO4 (Heptahydrate)
- 10 g EDTA*Na2 (Dihydrous)
- Solution #2 (500 mL ddH2O):
- 1.466 g CaCl2 (Dihydrate)
- 1.012 g MnCl2 (Tetrahydrate)
- 0.22 g (NH4)6Mo7O24 (Tetrahydrate)
- 0.314 g CuSO4 (Pentahydrate)
- 0.322 g CoCl2 (Hexahydrate)
- 0.998 g FeSO4 (Heptahydrous)
- Solution #1 (500 mL ddH2O):
- Procedure:
- Prepare solution #1 mixture (in a 1L bottle) according to these criteria:
- Add EDTA*Na2 slowly, stirring with a stir bar while you do so.
- After EDTA*Na2 has been well dissolved (little to no cloudiness), add ZnSO4 scoop by scoop.
- For every 3 scoops, add 200 uL of 10 M KOH. This solution must not be incredibly acidic in order for each component to be soluble.
- You should end with a pH of 6. It would be optimal to make this solution with a pH probe attached.
- Prepare solution #2 mixture (in a 1L bottle) and add a stir bar to assure that all components are dissolved well.
- Do not let this solution sit long before autoclaving. This solution should be light pink/orange by the time you combine it with solution #1.
- Before autoclaving, combine bottles into a 2L bottle.
- Autoclave as soon as possible or else things will precipitate!
MM1 liquid recipe
- Total volume: 2L (adjust accordingly)
- Solution #1 (1L ddH2O):
- 5.06 g K2HPO4 (Anhydrous)
- 5.18 g NaH2PO4 (Monohydrous)
- Solution #2 (1L ddH2O):
- 0.4 g MgSO4 (Heptahydrous)
- 1 g (NH4)2SO4 (Anhydrous)
- Solution #1 (1L ddH2O):
- Procedure:
- Mix constituents of solution #1 and #2 separately in 2L bottles.
- Currently considering the effects of mixing #1 & #2 before autoclaving.
- Organize tiny bottles to be filled with media post autoclave.
- Each bottle should contain ~150 mL of media.
- Autoclave large bottles and tiny bottles.
- Add 2 mL Visnaic trace elements into bottle #2.
- Pour equal amounts of solution #1 and #2 into the smaller bottles.
MM1A recipe
- Total volume: 2L (adjust accordingly)
- Solution #1 (1L ddH2O):
- 5.06 g K2HPO4 (Anhydrous)
- 5.18 g NaH2PO4 (Monohydrous)
- Solution #2 (1L ddH2O):
- 0.4 g MgSO4 (Heptahydrous)
- 1 g (NH4)2SO4 (Anhydrous)
- 30 g Bacto Agar
- Solution #1 (1L ddH2O):
- Procedure:
- Mix constituents of solution #1 and #2 separately in 2L bottles.
- Currently considering the effects of mixing #1 & #2 before autoclaving.
- Organize tiny bottles to be filled with media post autoclave.
- Each bottle should contain ~150 mL of media.
- Autoclave large bottles and tiny bottles.
- Add 2 mL Visnaic trace elements into bottle #2.
- Pour equal amounts of solution #1 and #2 into the smaller bottles.
- Let cool until solidification for storage.
- If you wish to pour plates immediately after autoclaving, place bottles into 50C incubator before pouring into plates.