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RNA extraction protocol

Arabidopsis RNA extraction protocol

Based on Krapp at al (1993) Plant Journal 3:817 with modifications.
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  1. 1-2 g fresh material, freezer-dried, ground with 0.2g sand (if necessary), and then homogenized with 10ml RNA extraction buffer (see below).
  2. Spin at 8,000rpm, 4oC, for 10 minutes
  3. Remove the supernatants to new tubes, phenol/chloroform extracted (8,000rpm at 4oC 10’).
  4. Wash the supernatant with 10 ml chloroform (8,000rpm at 4oC, 10’).
  5. Add 1/10 vol of 3M NaAc (700ul), and 2 vol of ethanol (15ml), -80oC 2hrs.
  6. Centrifuge at 8,500rpm for 30min at 4oC, and discard the supernatant.
  7. Resuspend the pellet in RNA resuspension buffer (see below), 4 oC 1hr.
  8. Centrifuge at 8,500rpm for 10min at 4oC, resuspend in RNase-free water (1 to 1.5ml).
  • RNA extraction buffer:
  • RNA resuspension buffer:

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