Plant Cell Reports

, Volume 15, Issue 7, pp 516–521

A reliable method for extraction of RNA from various conifer tissues

  • Jin-Zhuo Dong
  • David I. Dunstan
Article

Summary

A simple and efficient procedure suitable for extraction of high-quality RNA from cultured conifer tissues, somatic embryos, zygotic embryos, needles, stem and root tissues was developed. It produced from 100 μg up to 700 μg total RNA per gram tissue dependent on the types of tissues used. RNA quality was estimated by spectrophotometry, agarose gel electrophoresis, in vitro translation of mRNA, cDNA synthesis and Northern blot analysis. The method also worked well with Arabidopsis thaliana and tobacco tissues.

Key words

conifer Picea glauca RNA extraction 

Abbreviations

CTAB

cetyltrimethylammonium bromide

DEPC

diethylpyrocarbonate

PVP

polyvinylpyrrolidone

SDS-PAGE

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bahloul M, Burkard G (1993) Plant Mol. Biol. Repr. 11:212–215.Google Scholar
  2. Baker SM, White EE, Misra S (1991) Can. J. For. Res. 22:622–624.Google Scholar
  3. Baker SS, Rugh CL, Kamalay JC (1990) Biotechnique 9:268–272.Google Scholar
  4. Bommineni VR, Chibbar RN, Datla RSS, Tsang EWT (1993) Plant Cell Rep. 13:17–23.Google Scholar
  5. Chirgwin JM, Przybyla AE, MacDonald PJ, Rutter WJ (1979) Biochemistry 18:5294–5299.PubMedGoogle Scholar
  6. Dunstan DI, Bethune TD, Abrams SR (1991) Plant Sci. 76:219–228.Google Scholar
  7. Graham GC (1993) Plant Mol. Biol Repr. 11:32–37.Google Scholar
  8. Hughes DW, Galau G (1988) Plant Mol. Biol. Rep. 6:253–257.Google Scholar
  9. Jefferson RA, Kavanagh TA, Bevan MW (1987) EMBO J. 6:3901–3907.Google Scholar
  10. John ME (1992) Nuc. Acids Res. 20:2381.Google Scholar
  11. Lewinsohn E, Steele CL, Croteau R (1994) Plant Mol. Biol. Rep. 12:20–25.Google Scholar
  12. Logemann J, Schell J, Willmitzer L (1987) Anal. Biochem. 163:16–20.Google Scholar
  13. Maes M, Messens E (1992) Nuc. Acids Res. 20:4374.Google Scholar
  14. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor University Press, Cold Spring Harbor.Google Scholar
  15. Pelle R and Murphy NB (1993) Nuc. Acids Res. 21:2783–2784.Google Scholar
  16. Pich U, Schubert I (1993) Nuc. Acids Res. 21:3328.Google Scholar
  17. Shirzadegan M, Christie P, Seemann JR (1991) Nuc. Acids Res. 19:6055.Google Scholar
  18. Wadsworth GJ, Redinbaugh MG, Scandalios JG (1988) Anal. Biochem. 172:279–283.Google Scholar
  19. Whitmore FW, Knebel HB (1987) Can J. For. Res. 17:408–412.Google Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Jin-Zhuo Dong
    • 1
  • David I. Dunstan
    • 1
  1. 1.Plant Biotechnology Institute, National Research Council of CanadaSaskatoonCanada

Personalised recommendations