RNA isolation using phase extraction and LiCl precipitation View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2005

AUTHORS

Catherine Kistner , Manuel Matamoros

ABSTRACT

The extraction of good quality RNA from plant tissues is a critical step in gene expression studies. We have developed a protocol for RNA isolation from Lotus japonicus roots and leaves that yields RNA that can be successfully used in downstream applications such as reverse transcription-polymerase chain reaction (RT-PCR). The use of an adequate extraction buffer is crucial to obtaining good quality RNA, especially when working with roots. The composition of the extraction buffer has been optimized to avoid contamination with phenolic compounds, which are very abundant in Lotus roots and can inhibit downstream enzymatic reaction such as reverse transcription. In our laboratory, this method yields highly purified RNA suitable for cDNA synthesis and is routinely used for gene expression studies in L. japonicus roots. For Northern blots, dirtier (i.e. phenolic contamination), higher yielding extraction methods might be suitable as well. More... »

PAGES

123-124

Book

TITLE

Lotus japonicus Handbook

ISBN

1-4020-3734-1

Author Affiliations

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/1-4020-3735-x_9

DOI

http://dx.doi.org/10.1007/1-4020-3735-x_9

DIMENSIONS

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