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Figure 3 | Plant Methods

Figure 3

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

Figure 3

Consistency and reproducibility of HTP plant RNA extraction procedure. A: The HTP RNA procedure was performed manually using multi-channel pipettes with either centrifugation or vacuum to pass liquids through the binding plate or automated using a liquid handling robot with a vacuum. All three protocols (manual centrifuge, manual vacuum and automated vacuum) isolated similar amounts of RNA (loading lysate from ~20 mg plant tissue per well). Data are expressed as mean RNA concentration (ng/μl) per well of 96 well plate ± SD. B: Results from 4 different 96 well plates processed using the automated procedure (384 samples) demonstrate that the method is reproducible both intra- and inter-plates. Results are presented as a box and whisker plot of the concentration of RNA isolated (ng/μl; each well containing 50 μl). The bottom and top of the boxes are the 25th and 75th percentiles and the band in the middle is the median. The ends of the whiskers are 1.5 × interquartile range and data points outside this are considered outliers. There are no outliers in plate 1, 1 upper (230 ng/μl) and 2 lower (87 ng/μl and 96 ng/μl) in plate 2, 1 lower outlier (56 ng/μl) in plate 3 and 2 lower outliers (86 ng/μl and 87 ng/μl) in plate 4. The maximum and minimum outliers for each plate (if any) are shown in the box and whisker plot.

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