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

Content Provider	Springer Nature Link
Author	Xu, Qiang Wen, Xiaopeng Deng, Xiuxin
Copyright Year	2004
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.
Starting Page	301
Ending Page	302
Page Count	2
File Format	PDF
ISSN	07359640
Journal	Plant Molecular Biology Reporter
Volume Number	22
Issue Number	3
e-ISSN	15729818
Language	English
Publisher	Springer-Verlag
Publisher Date	2012-09-03
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	chestnut rose (Rosa roxburghii Tratt) DNA isolation PCR RFLP polysaccharides Plant Sciences Plant Breeding/Biotechnology Proteomics Metabolomics Bioinformatics
Content Type	Text
Resource Type	Article
Subject	Plant Science Molecular Biology

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