Evaluation and Optimization of Genomic DNA Isolation Protocols from Human Solid Tissues

Abstract

Molecular diagnostics are performed by using DNA isolated from different body tissues. However, it is necessary to obtain genomic DNA of good quality. The objective of this study was to describe the efficient protocol of DNA extraction from human breast, adipose, brain, liver, kidney, prostate, lung, larynx, endometrium and muscle tissues. We obtained high molecular weight DNA of good quality, shown by agarose gel and DNA fragments. Spectrophotometric analysis of DNA concentration showed variation among the DNA from different tissues, with the liver, breast, endometrium and adipose tissues presenting the greatest and the smallest concentration, respectively. The described protocol has proven to be advantageous due to its simplicity, quickness, affordable reagents and absence of phenol, resulting in a high molecular weight DNA of good quality from several tissues. Genomic DNA extraction from solid tissues of patients with cancer risks was carried out by using two different procedure including manuel and automatically isolation. The QIAamp spin columns (QIAGEN, Hilden, Germany) were used for extraction and purification of genomic DNA from different tissues by Tissue Kits and Tissue Card or automatically, EZ1 automatic DNA isolation system. Furthermore, we used Phenol-Chloroform protocol for manuel isolation in this study

Keywords

• nDNA, solid tissue