I want to estimate the total genome size of my study plants. These plants are found in the high Himalayas. Are there any reliable, simple protocols for the estimation of total genome size of plant from leaf material?
Are there any lab in India working on plant total genome size estimation by flow cytometry??
I would suggest you to try out Otto's buffer. This can earn you a good result.
Otto I buffer (Otto, 1990): 100 mM Citric Acid; 0.5 % (v/v) Tween 20 (pH approx. 2.3)
Otto II buffer (Doležel and Göhde, 1995): 400 mM Na 2 HPO 4 .12H 2 O (pH approx 8.9)
Take around 50 mg leaf tissue without midrib, chop it in 500 ul of Otto I buffer. Add 100 ug/ ml conc of PI in 500 ul Otto II buffer. Mix it with 500 ul of nuclei suspension in Otto I. Pass the nuclei suspension through a 50 micron nylon mesh, run in FCM.
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