纯化基因组DNA，使蛋白质污染降至最低。
Purification of genomic DNA with minimal contamination of proteins.
作者信息
Hebron Haroun R, Yang Yu, Hang Jun
机构信息
EdgeBio, Gaithersburg, Maryland 20877, USA.
出版信息
J Biomol Tech. 2009 Dec;20(5):278-81.
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.
该纯化方法基于一组溶液和简单的离心程序。设计的方案可轻松从多种样本中提取和纯化基因组DNA，这些样本包括全血、血沉棕黄层、骨髓、体液、颊细胞、组织、小鼠尾巴等。通过稀释到低渗溶液中来裂解红细胞。在阴离子去污剂存在的情况下，用蛋白酶K分解并消化组织以释放基因组DNA。在去污剂和蛋白质沉淀后，加入能结合蛋白质、脂质和RNA的独特磁珠以实现最高纯度。然后通过乙醇沉淀分离基因组DNA。使用一种专利核酸沉淀试剂来提高从低浓度样本中回收DNA的效率。该方法不使用DNA结合磁珠或柱，消除了产量低的问题以及基因组DNA剪切的风险。纯化后的样本不含蛋白质、脂质、盐和RNA污染。纯化后的样本储存也很稳定，适用于所有下游应用。
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