I want to do ABA treatment in Arabidopsis thalliana. I am looking for some protocols in which I can dip the sample in ABA and then evaluate for some gene expression after some hours.
Hi Chirag,
There are various ways you can treat your AT plants with ABA. As yous aid you want to check gene expression, I will tell you the simplest strategy.
Start with seed sterilization, store sterilized seeds in 4 degree refrigerator for 3-4 days.
After 4 days sow seeds on MS agar plates (try to keep concentration of agar little low than usual as later you will need to pull out plants) and keep the plates in suitable light, temperature and humidity conditions.
Once the plants are 7-8 days old, take them out ( I will take out approx 20 plants) and place them in a petridish with a whatman paper soaked with MS medium + Sucrose (No agar) and different concentrations of ABA. Keep plates with your saplings under proper light/temperature/humidity conditions.
Have at least 3 plates for same concentration of ABA.
Decide how long you want to keep your plants soaked in ABA and after that you can extract RNA and go for checking gene expression.
Good luck.
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