Literature DB >> 19949702

Purification of genomic DNA with minimal contamination of proteins.

Haroun R Hebron1, Yu Yang, Jun Hang.   

Abstract

The purification is based on a set of solutions and a simple centrifugation procedure. Protocols are designed for an easy extraction and purification of genomic DNA from a wide range of samples, including whole blood, buffy coat, bone marrow, body fluids, buccal cells, tissues, mouse tails, etc. RBCs are lysed by dilution into a hypotonic solution. Tissues are broken down and digested by proteinase K in the presence of an anion detergent to release genomic DNA. After precipitation of the detergent and proteins, unique beads that bind proteins, lipids, and RNAs are added to achieve the supreme purity. Genomic DNA is then separated by alcohol precipitation. A proprietary nucleic acid precipitation reagent is used to enhance DNA recovery from low concentration samples. No DNA-binding beads or columns are used in the method, eliminating the problem of low yield and the risk of shearing of genomic DNA. The purified samples are free of proteins, lipids, salts, and RNA contamination. Purified samples are also stable for storage and suitable for all downstream applications.

Entities:  

Keywords:  blood and tissue; ethanol precipitation; nucleic acid purification

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Substances:

Year:  2009        PMID: 19949702      PMCID: PMC2777350     

Source DB:  PubMed          Journal:  J Biomol Tech        ISSN: 1524-0215


  3 in total

1.  Development of a Membrane-Based Method for Isolation of Genomic DNA from Human Blood.

Authors:  Goutam Mandal; Subhasish Das; Sriram Padmanabhan
Journal:  J Biomol Tech       Date:  2018-03-23

2.  TENT: A Rapid DNA Extraction Method of Staphylococcus aureus.

Authors:  Sepideh Hassanzadeh; Mohammad Reza Pourmand; Davoud Afshar; Sanaz Dehbashi; Rahil Mashhadi
Journal:  Iran J Public Health       Date:  2016-08       Impact factor: 1.429

3.  pH-dependent sedimentation of DNA in the presence of divalent, but not monovalent, metal ions.

Authors:  Corbin J England; Tanner C Gray; Shubha R L Malla; Samantha A Oliveira; Benjamin R Martin; Gary W Beall; L Kevin Lewis
Journal:  Anal Biochem       Date:  2020-12-31       Impact factor: 3.365

  3 in total

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