Hi everyone,
I work with Arabidopsis thaliana.
I want to know what is the best strategy to strore plant saplings after the treatment to preserve the quality of RNA (at least for a week).
1. Is it adviseable to flash freeze sapling in a microcetrifuge tube in liquid nitrogen and keep it in -80
or
2. Is it better to add RNA lysis buffer, crush the samples and then flash freeze and keep it in -80.
or
3. I should just add the RNA lysis buffer without crushing the saplings?
I know if I flash freeze the saplings without any buffer the ice crystals will still puncture the plant cell, might lead the slow yet possile RNAse activity thereby RNA damage, wherease I think adding RNA lysis buffer will act as inhibitior of that slow RNAse activity during the storage period.
Any other suggestions are most welcome.
I will be happy to get other useful advices that work in your case.
thanks
You can try using RNA later solution, it helps preserve the RNA even in fresh samples (i.e., unfrozen samples). It has worked with my animal tissue. Here is the link to purchase the product-
https://www.thermofisher.com/order/catalog/product/AM7021
Hope it helps.
Crush the plant samples in RNA lysis buffer, freeze them in liquid nitrogen, and keep the vial at -80 degree centigrade. RNase inhibitors are usually present in the RNA lysis buffer, preventing RNA from degrading.
But it is better to use RNAlater solution for plant material preservation because it contains a high amount of quaternary ammonium sulfates and cesium sulfates which denature RNases and prevent RNA degradation.
Flash-freezing in liquid nitrogen is the best. Even better, grind them liquid nitrogen to a fine powder and store the powder in -80C.
Plant RNA kits all start with needing to grind your samples to disrupt.
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