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Figure 2 | Plant Methods

Figure 2

From: Protocol: high throughput silica-based purification of RNA from Arabidopsis seedlings in a 96-well format

Figure 2

Detailed overview of the HTP plant RNA extraction procedure. The HTP total RNA extraction procedure processes 96 samples simultaneously using the illustra RNAspin 96 RNA isolation kit. A: Seedlings are collected directly into RNAlater solution to preserve RNA integrity. B: Plant tissue is homogenised using a bead mill. C: Lysed samples are filtered to remove debris and half stored in case of catastrophe. RNA is extracted from the other half. D: RNA is prepared for binding to the silica membrane. E: RNA binds to the silica membrane and unbound material is removed by a series of buffer washes. F: Purified RNA is eluted by centrifugation. Sections D and E (boxed) can be performed manually using multichannel pipettes and either centrifugation or vacuum to pass solutions through the binding plate or alternatively automated using a liquid handling robot with a vacuum manifold. The approximate time taken for each step is indicated on the right. Although the automated section is only slightly quicker in terms of actual time taken to complete the procedure, the automated protocol does not require any hands on intervention of the researcher thus freeing up time to complete other work. These factors combine resulting in a 4 fold increase in possible throughput using the automated rather than the manual protocol. Images of the illustra RNAspin 96 kit plates © 2011 General Electric Company - reproduced by permission of the owner.

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