Arabidopsis RNA extraction protocol

Based on Krapp at al (1993) Plant Journal 3:817 with modifications.

Comments or Questions? Send to me at: chunming.liu@wur.nl

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1-2 g fresh material, freezer-dried, ground with 0.2g sand (if necessary), and then homogenized with 10ml RNA extraction buffer (see below).

Spin at 8,000rpm, 4oC, for 10 minutes

Remove the supernatants to new tubes, phenol/chloroform extracted (8,000rpm at 4oC 10’).

Wash the supernatant with 10 ml chloroform (8,000rpm at 4oC, 10’).

Add 1/10 vol of 3M NaAc (700ul), and 2 vol of ethanol (15ml), -80oC 2hrs.

Centrifuge at 8,500rpm for 30min at 4oC, and discard the supernatant.

Resuspend the pellet in RNA resuspension buffer (see below), 4 oC 1hr.

Centrifuge at 8,500rpm for 10min at 4oC, resuspend in RNase-free water (1 to 1.5ml).

Solutions: