Extraction of total RNA from leaves of Eucalyptus and other woody and herbaceous plants using sodium isoascorbate.

نویسندگان

  • Y Suzuki
  • T Hibino
  • T Kawazu
  • T Wada
  • T Kihara
  • H Koyama
چکیده

Rapid extraction of total RNA from Eucalyptus leaves is difficult due to the high content of polyphenolics and polysaccharides. A rapid and simple method was developed by using an extraction buffer containing sodium isoascorbate at a concentration of 500 mM. This method consisted of one or two chloroform extractions, one acid guanidium-phenol-chloroform extraction, and isopropanol precipitation alone. The yields of the RNA fractions were 246-1750 micrograms/g fresh weight when leaves of Eucalyptus, five other woody plants, and four herbaceous plants were used as samples. The contamination of the RNA fractions by proteins and polysaccharides was very limited as judged spectrophotometrically. When the RNA fractions were subjected to agarose gel electrophoresis, intact rRNA bands were detected. The RNA fractions could be used for RT-PCR. These results indicate that our new method achieves a simple and rapid preparation of high-quality RNA from leaves of Eucalyptus and other plant species.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Isolation of high-quality RNA from a wide range of woody plants

Isolation of high-quality RNA is one of the most crucial methods in molecular biology. RNA extraction from woody plants has been problematic due to the presence of rigid and woody tissues, large amounts of polysaccharides, polyphenols and other secondary metabolites. Here we present a suitable protocol for RNA isolation from a wide range of woody plants that includes eight gymnosperms and four ...

متن کامل

Volatile Oil Constituents of the Eucalyptus viridis R. T. Baker and Eucalyptus oleosa F. Muell. Leaves from Iran

  Background: The genus Eucalyptus (family Myrtaceae) comprises well-known plants of over 600 species of trees. Although most of the plants are native to Australia, numerous species have been introduced to other parts of the world, including Iran, as economic and ornamental trees in forest trial provenances, where the plants have become source of important fast-growing hardwood trees and Eucaly...

متن کامل

Multigene phylogeny and morphotaxonony of Septoria spp. from Iran along with a checklist of septoria-like taxa

The genus Septoria includes important plant pathogens with worldwide distribution, commonly associated with leaf spots and stem cankers of a broad range of plant hosts. In this study, eight isolates of Septoria were recovered from leaves with leaf spot on four herbaceous and woody plants from Gilan, Ardebil, East and West Azerbaijan provinces in north and northwest of Iran. The isolates were st...

متن کامل

Chloroplast DNA extraction from herbaceous and woody plants for direct restriction fragment length polymorphism analysis.

The technique described here is a fast and simple method of extracting chloroplast DNA (cpDNA). It overcomes the need for differential centrifugation using density gradients. The leaves do not have to be kept in the dark and lyophilized before extraction, but lyophilization is still possible. The chloroplasts are specifically lysed in a cell extract of leaves, using a non-ionic detergent. After...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • BioTechniques

دوره 34 5  شماره 

صفحات  -

تاریخ انتشار 2003