Plant Molecular Biology Reporter

, Volume 22, Issue 3, pp 269–278

High-quality RNA, cDNA, and derived EST libraries from grapevine (Vitis vinifera L.)

  • A. B. Iandolino
  • F. Goes da Silva
  • H. Lim
  • H. Choi
  • L. E. Williams
  • D. R. Cook
Genetic Resources

DOI: 10.1007/BF02773137

Cite this article as:
Iandolino, A.B., Goes da Silva, F., Lim, H. et al. Plant Mol Biol Rep (2004) 22: 269. doi:10.1007/BF02773137

Abstract

The implementation and generation of unbiased results from RNA-based techniques in functional genomic studies requires the isolation of high-quality RNA. In grapevines, the yield and quality of RNA can be significantly reduced by contaminants such as polyphenols, polysaccharides, and proteins, which are abundant during different stages of berry and leaf development. We describe an optimized cetyltrimethylammonium bromide (CTAB)-based RNA extraction protocol that allows effective extraction of high-quality total RNA from a wide range of grapevine tissues, including difficult organs such as symptomatic, pathogen-infected leaves and mature berries. Total RNA extracted with this protocol was successfully used for cDNA library construction as well as reverse transcription-polymerase chain reaction (RT-PCR) amplification. This protocol can also be easily scaled to accommodate different amounts of tissue.

Key words

CTABgrapevinepolyphenolspolysaccharidesRNA extractionVitis vinifera

Abbreviations

CTAB

cetyltrimethylammonium bromide

DEPC

diethyl pyrocarbonate

EtBr

ethidium bromide

PVP

polyvinylpyrrolidone

RT-PCR

reverse transcription-polymerase chain reaction

Copyright information

© International Society for Plant Molecular Biology 2004

Authors and Affiliations

  • A. B. Iandolino
    • 1
  • F. Goes da Silva
    • 2
  • H. Lim
    • 2
  • H. Choi
    • 2
  • L. E. Williams
    • 1
  • D. R. Cook
    • 2
  1. 1.Department of Viticulture and EnologyUniversity of California, DavisDavisUSA
  2. 2.Department of Plant PathologyUniversity of California, DavisDavisUSA