Mandal Goutam, Das Subhasish, Padmanabhan Sriram
Cancyte Technologies Pvt. Ltd., Sri Shankara Research Center, Rangadore Memorial Hospital, Bangalore 560004, India.
J Biomol Tech. 2018 Jul;29(2):46-53. doi: 10.7171/jbt.18-2902-001. Epub 2018 Mar 23.
High quality and sufficient quantity of genomic DNA (gDNA) are the primary requisites of several molecular biologic applications, including clinical studies related to genetics, genomics, gene polymorphism, and DNA fingerprinting. Whole blood is the primary source of gDNA in most of the clinical investigations. Currently, commercial kits are primarily used to achieve these goals. However, the use of kits is limited by the cost and involvement of several centrifugal steps. Other methods reported are either laborious or do not produce high quality or quantity of gDNA or both. Here, we present the data on the development of a centrifugation-free, cost-effective, and user-friendly method for the isolation of human gDNA from the buffy coat of human blood that involves limited numbers of steps with about 15 min of hands-on time per sample.
高质量和足够数量的基因组DNA(gDNA)是多种分子生物学应用的首要条件,包括与遗传学、基因组学、基因多态性和DNA指纹识别相关的临床研究。在大多数临床研究中,全血是gDNA的主要来源。目前,主要使用商业试剂盒来实现这些目标。然而,试剂盒的使用受到成本和多个离心步骤的限制。报道的其他方法要么费力,要么无法产生高质量或高数量的gDNA,或者两者都无法实现。在此,我们展示了一种从人血的血沉棕黄层中分离人gDNA的无离心、经济高效且用户友好的方法的开发数据,该方法步骤有限,每个样本的实际操作时间约为15分钟。