Arabidopsis Pollen Germination Medium (Leonor 2007)
There are a
For your convenience, we have given the molecular weights of each reagent and for its most used hydrated forms too. It is up to the user to determine whether hydrated forms make any difference for the experiments or not.
The pH of the medium should be adjusted to 7.5 using dilute HCl and KOH.
|Reagent||M.W||Working concentration||Stock solution||Add for 10 ml stock||Add from stock for 1X of 50ml|
|H2BO3||61.8||0.01%||1%||0.1 g||0.5 ml|
|CaCl2(anhydrous)||110.98||5mM||1M||1.1 g||250 µl|
|CaCl2 .2H2O||147.01||5mM||1M||1.4 g||250 µl|
|KCl||74.5||5mM||1M||0.745 g||250 µl|
|MgSO4||120.36||1mM||1M||1.2 g||50 µl|
|MgSO4 .7H2O||246.47||1mM||1M||2.465 g||50 µl|
|Noble Agar||-||1.50%||-||-||0.75 g|
- Boric acid
- Calcium chloride
- Potassium chloride
- Magnesium sulphate
- Noble agar
- Distilled water
Instruments and other requirements
- Weighing balance. paper boats.
- Graduated cylinder and a glass beaker.
- pH meter. Magnetic stirrer and magnetic bead/pellet.
- Tissue paper.
Use ddH2O only for making PGM
You can choose to make 1X (working concentration) or make stocks and use them to make 1X whenever you want. Making stock solutions eliminate the possibility of errors that may happen while measuring very low quantities of reagents. Making working concentration from stocks also helps you to add some other reagents without further diluting working concentration.
- Determine whether you want to start with stock solutions or directly making working concentration.
- Weigh the reagents accordingly (Refer to the table above).
- Dissolve salts by using magnetic stirrer or vortex mixer.
- Adjust pH to 7.5 using HCl and KOH (pH is one of the critical factors)
- Make the volume as planned.
- Sterilization of media is not required for immediate use. However, doing filter sterilization might increase its shelf life by preventing microbial growth.
- Aliquot to smaller volumes if needed (frequent opening and closing in open air might result in microbial growth).
- Noble agar is preferable over agar as solidifying agent.
Store PGM (Pollen Germination Medium) stock solutions at 4˚C for 1-2 weeks.
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