RNA isolation from plant cells

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RNA isolation from plant cells

by Chris Forreiter, adapted from Planta 190 (1993) 536-45

- put cells in GTC buffer
- 10' 37 C
- precipitate RNA o/n @ -20 C with 2 vol. 3M NaOAc and 0.75 vol. EtOH
- 15' 14000 rpm @ 4 C
- pellet in TES (TE + 0.1% SDS)

optional:
- phenol/chloroform extraction
- remove dsDNA by precipitation of RNA with 1/3 vol. 8M LiCl, 2h @ 4 C (NO EtOH or iPrOH). pellet in TE, then normal EtOH precipitation

GTC buffer

4.5 M GTC (guanidinium isothiocyanate)
50 mM HEPES/KOH pH 7.5
2 % lauroylsarcosine
1 % b-mercaptoethanol

filter, DO NOT AUTOCLAVE

10 mM Tris pH 8, 1 mM EDTA

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