Genomic DNA extraction fromVictoria amazonica

DNA extraction is difficult in many plants because of metabolites that interfere with DNA isolation procedures and subsequent applications such as DNA restriction, amplification, and cloning. We developed the first reliable and efficient method for isolatingVictoria amazonica genomic DNA that is fre...

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Bibliographic Details
Published in:Plant molecular biology reporter 2004-06, Vol.22 (2), p.195-196
Main Authors: Puchooa, Daneshwar, Khoyratty, Sher-Ullah Shahnoo Sulliman
Format: Article
Language:English
Subjects:
Cloning
Deoxyribonucleic acid
DNA
Genomics
High performance liquid chromatography
Liquid chromatography
Metabolites
Polymorphism
Polyphenols
Polysaccharides
Polyvinylpyrrolidone
Saccharides
Sodium
Sodium chloride
Sodium sulfite
Spectrophotometry
Sulfite
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Summary:DNA extraction is difficult in many plants because of metabolites that interfere with DNA isolation procedures and subsequent applications such as DNA restriction, amplification, and cloning. We developed the first reliable and efficient method for isolatingVictoria amazonica genomic DNA that is free from polysaccharides and polyphenols. This protocol uses 1.5 M NaCl, 2% polyvinylpyrrolidone (PVP) (Mr 1000), 5% mercaptoethanol, 0.12% sodium sulfite, and an incubation at 65°C for 4 h. The purity of isolated genomic DNA was confirmed by means of high-performance liquid chromatography (HPLC) profile and spectrophotometric analyses (A260/230 ratio of 1.836, A260/280 of 1.842). DNA was obtained in the amount of 387 μg per gram of leaf material, and it proved amenable to restriction digestion.
ISSN:0735-9640
1572-9818
DOI:10.1007/BF02772727