Can anyone kindly tell me what amount of rice seedling leaves and root is required for isolation of good quantity of RNA ? Should I weigh the freshly chopped samples before homogenizing with lN2 or after?
Also can anyone please suggest methods for storage of rice seedling root and shoot samples for later RNA isolation?
Take a rough leaf and measure the amount for 100mg(roughly) According to that you can have a approx idea that how much tissue you required for 100mg sample,then cut the leaves you want to take.After cutting the leaves, wipe it with RNAseZAP(I used Invitrogen), then directly proceed for RNA isolation by DEPC treated Mortar Pestle(Homogenization).
All the best.
There is no any fixed amount. Normally 100-120mg is enough for RNA extraction. Don't worry about the amount. As Dipanwita Datta said you can assume the amount. After collection immediately put into liquid nitrogen and keep it until you start extraction.
For long term storage RNA Later/Trizol can be used. In my case, I stored samples at -70oC with Trizol.
We recently isolated RNA from 40 mg rice blade tissue and got >6 ug/ul using RNeasy kit. Using young, healthy tissue is the key.
Thank you everyone for the information.
@Dipanwita Datta should i wipe the leaves individually with RNAase ZAP or wearing a glove while cutting the leaves using a sterile blade should do?
Hello Anitha,
Wear gloves, cut the whole leaf, wipe with RNAse Zap and then cut into pieces and dip it in liquid nitrogen but Do it fast and dip it in Liquid nitrogen.
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