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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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Published in: | Plant molecular biology reporter 2004-06, Vol.22 (2), p.195-196 |
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container_title | Plant molecular biology reporter |
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creator | Puchooa, Daneshwar Khoyratty, Sher-Ullah Shahnoo Sulliman |
description | 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. |
doi_str_mv | 10.1007/BF02772727 |
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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 |
title | Genomic DNA extraction fromVictoria amazonica |
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