Plant Molecular Biology Reporter

, Volume 22, Issue 3, pp 301–302

A simple protocol for isolating genomic DNA from chestnut rose (Rosa roxburghii tratt) for RFLP and PCR analyses

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DOI: 10.1007/BF02773140

Cite this article as:
Xu, Q., Wen, X. & Deng, X. Plant Mol Biol Rep (2004) 22: 301. doi:10.1007/BF02773140

Abstract

Isolation of high-quality DNA from rosaceous species is particularly difficult because of their high levels of polyphenols, polysaccharides, and other compounds. The yields and quality of genomic DNA are considerably affected when the common protocol for DNA isolation is applied to the chestnut rose (Rosa roxburghii Tratt). A simple, rapid, and efficient protocol for the extraction of DNA from the chestnut rose is described. The modified hexadecyltrimethylammonium bromide (CTAB) procedure, which uses phenol-absent extraction to enhance the yield, involves a washing step before extraction for the removal of organic molecules and excessive water; the use of high concentrations of polyvinylpyrrolidone (2% [w/v]), CTAB (3% [w/v]), and β-mercaptoethanol (3% [v/v]) in the high-salt-concentration extraction buffer to remove polyphenols and polysaccharides; and the combined use of potassium acetate and chloroform to remove proteins and polysaccharides. Finally, DNA is precipitated with an equal volume of isopropanol and 0.1 vol of sodium acetate. This protocol results in high yields of DNA. The average yield of DNA ranged from 980–1800 μg/g of fresh weight of leaves. Downstream results indicate that DNA quality is sufficient for restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR) analyses.

Key words

chestnut rose (Rosa roxburghii Tratt)DNA isolationPCRRFLPpolysaccharides

Copyright information

© International Society for Plant Molecular Biology 2004

Authors and Affiliations

  1. 1.National Key Laboratory of Crop Genetic ImprovementHuazhong Agriculture UniversityWuhanPR China
  2. 2.Guizhou Key Laboratory of Agricultural BiotechnologyGuizhou UniversityGuiyangPR China