RNA purification

Total RNA is extracted from plants using TRIzol reagent according to the supplier's instructions (Takara, Japan, or equivalent). For homogenizing plant material, plants and four steel balls are placed in a 2-ml Eppendorf tube and homogenized using a Shake Master version 1.2 (BMS-12). Alternatively, RNA can be extracted from plants using the ATA method. However, because ATA inhibits the activity of reverse transcriptase, the RNA cannot be used for real-time polymerase chain reaction (RT-PCR) and cDNA synthesis. For a detailed protocol of this method, see Nakashima and Yamaguchi-Shinozaki (2002).

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