Protocol

RNA Extraction from Arabidopsis for Northern Blots and Reverse Transcriptase-PCR

Adapted from Arabidopsis: A Laboratory Manual (eds. Weigel and Glazebrook). CSHL Press, Cold Spring Harbor, NY, USA, 2002.

INTRODUCTION

Analysis of the cellular distribution of gene products (RNA and protein) can provide important clues to the function of genes in vivo. RNA expression can be monitored either after extraction from plants or tissues or in situ. The former procedure is easier to perform and more quantitative, whereas the latter provides much more detailed resolution of spatial and temporal expression patterns but is often more technically challenging. This protocol describes a method for RNA extraction from Arabidopsis thaliana. Frozen plant tissue is homogenized using a hand drill fitted with a plastic micropestle instead of using a conventional mortar and pestle, which can be inefficient and physically demanding. This homogenization method allows samples to be processed quickly (up to 12 samples/hour). Typical yields are 20-40 µg of total RNA from 100 mg of Arabidopsis seedlings. The RNA extracted using this method is suitable for reverse transcriptase-polymerase chain reaction (RT-PCR) and Northern hybridization.