RNA extraction from arabidopsis for northern blots and reversetranscriptase-PCR

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

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.

Original languageEnglish
JournalCold Spring Harbor Protocols
Volume4
Issue number9
DOIs
Publication statusPublished - 2009 Dec 7

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Arabidopsis
Northern Blotting
RNA
Polymerase Chain Reaction
Tissue
Homogenization method
Mandrillus
Polymerase chain reaction
RNA-Directed DNA Polymerase
Reverse Transcriptase Polymerase Chain Reaction
Mortar
Seedlings
Plastics
Hand
Genes
Proteins

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

RNA extraction from arabidopsis for northern blots and reversetranscriptase-PCR. / Ahn, Ji H.

In: Cold Spring Harbor Protocols, Vol. 4, No. 9, 07.12.2009.

Research output: Contribution to journalArticle

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