Triturating plant tissue before isolation

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Mar 14, 2017
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Hello,

It's my first time in this place, because I've just started my adventure with molecular aspects of biology. So please, be tolerant

In last time I isolated plant tissue (roots, leaves), however the amount, which I got, was very low. Before isolation, I triturated plants' organs in liquid nitrogen. After that I did the isolation with Tri-reagent (Sigma) according the protocol. First time, when I got small account of RNA (200-300 [ng/µl]), I changed reagents (I used new isoprophanol, ethanol, DEPC), but the problem didn't disappear. So, is it possible, that too many/too little amount of the liquid nitrogen, which I used during triturating organs, is the reason of my problem?

Other hand - I had to use organs from Cucumis sativus (Krak) - plants grew three weeks on full-medium, however during trird week on the leaves appeared chlorosis and nerosis. If it may be a reason of so low account of RNA from roots and leaves, which I isolated?

I will be very thankful for answers and suggests!

1 Comments

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nekameneka 10 months ago

Hi wnt,

Sorry for late reply, I hope you still read it. I would like to know why you use nitrogen to do that? I think you lost a lot of RNA in this process.