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Research Paper | Biology | India | Volume 4 Issue 11, November 2015 | Popularity: 6.7 / 10

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Isolation of High Quality DNA from Pulverized Rice Grain Samples by Tri-Phase Method

Jithin Thomas, V. J. Dominic

Abstract: Genetic studies in plants require the isolation of high quality DNA from large number of plant sources. Commercial DNA extraction kits are convenient and usually safe, but their availability to certain developing countries and high cost of available kits can be limiting factors, especially when handling a large number of samples and considering experiments with limited financial resources. There has been a need for reliable, inexpensive and safe method to isolate DNA from milled/pulverized rice samples. Hence, a simple, economical and reliable method is presented to isolate high quality DNA from milled rice samples. DNA isolation is conducted in three phases, cell lysis phase, DNA precipitation phase and de-proteinizing phase. Initially cell lysis is done by extraction buffer, later K+ ions in Potassium acetate helps in replacing Na+ ions. Hence, dodecyl sulphate coated starch and protein complexes become precipitated. Further DNA purification is done by de-proteinizing treatment with phenol chloroform ethanol, mixture. DNA obtained by this method can be subjected to enzymatic reactions, such as digestion with restriction enzymes, ligation and PCR amplification.

Keywords: SDS sodium dodecyl sulphate, genomic DNA, Molecular weight MW, Polymerase Chain Reaction PCR

Edition: Volume 4 Issue 11, November 2015

Pages: 524 - 527

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