Hostname: page-component-78c5997874-g7gxr Total loading time: 0 Render date: 2024-11-08T02:38:36.342Z Has data issue: false hasContentIssue false

Rapid and efficient purification of seed messenger RNA without phenol:chloroform extraction

Published online by Cambridge University Press:  19 September 2008

S. K. Pramanik*
Affiliation:
Department of Botany, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
T. L. Reynolds
Affiliation:
Department of Botany, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
S. A. Maclsaac
Affiliation:
Department of Botany, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
J. D. Bewley
Affiliation:
Department of Botany, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
*
* Correspondence

Abstract

A novel method has been devised to extract poly (A+)RNA from seed tissues. The advantages of this technique include the speed of extraction and the elimination of the hazardous phenol:chloroform extraction step which usually precedes oligo (dT)-cellulose column chromatography. Instead, the crude supernatant, containing a high concentration of SDS and salts to effectively inhibit RNase activity, is passed directly over the oligo (dT)-cellulose column. The resultant isolated poly (A+)RNA is of high yield and purity and is suitable for in vitro translation and Northern blot analysis.

Type
Short Communication
Copyright
Copyright © Cambridge University Press 1993

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Casey, R., Domoney, C. and Ellis, N. (1986) Legume storage proteins and their genes. Oxford Surveys of Plant Molecular and Cell Biology 3, 295.Google Scholar
Krochko, J.E., Pramanik, S.K. and Bewley, J.D. (1992) Contrasting storage protein synthesis and messenger RNA accumulation during development of zygotic and somatic embryos of alfalfa (Medicago sativa L.). Plant Physiology 99, 4653.CrossRefGoogle ScholarPubMed
Pramanik, S.K., Krochko, J.E. and Bewley, J.D. (1992) Distribution of cytosolic mRNAs between polysomal and ribonucleoprotein complex fractions in alfalfa embryos. Stage-specific translational repression of storage protein synthesis during early somatic embryo development. Plant Physiology 99, 15901596.CrossRefGoogle ScholarPubMed
Pramanik, S.K., Walsh, R.W. and Bag, J. (1986) Association of mRNA with the cytoskeletal framework in rat L-6 myogenic cells. European Journal of Biochemistry 160, 221229.CrossRefGoogle Scholar
Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular cloning: a laboratory manual. Cold Spring Harbour, New York, Cold Spring Harbour Laboratory.Google Scholar