RNA Extraction for Northern Blots and RTPCR

This protocol was contributed by ]i Hoon Ahn (Salk Institute).

There are several kits available for RNA extraction, e.g., RNeasy (QIA-GEN). However the cost of these kits can be prohibitive for processing a large number of samples. The following protocol is based on a method described by Chomczynski and Sacchi (1987). Instead of using a conventional mortar and pestle, which can be inefficient and physically demanding, frozen plant tissue is homogenized using a hand drill fitted with a plastic micropesde. This homogenization method allows samples to be processed more quickly (up to 12 samples/hour). Typical yields are 20-40 fig of total RNA from 100 mg of Arabidopsis seedlings. The total RNAs are suitable for reverse transcriptase-polymerase chain reaction (RT-PCR) and northern hybridization.

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