Phenol-Chloroform DNA Isolation Method

Isolation of DNA through phenol-chloroform is one of the most commonly used methods in molecular biology. Researchers can obtain a good yield of high molecular weight DNA from any biological sample using a mixture of phenol-chloroform-isoamyl alcohol. The basic principle involves the separation of DNA, RNA, and proteins in a sample, based on the differential solubilities of these molecules in different immiscible liquids. The isolated DNA can be used in gene amplification, fingerprinting, single nucleotide polymorphism studies, RFLP, and gene quantification. Though the present chapter describes the isolation of DNA from blood cells, the basic steps are similar for DNA isolation from other tissues or cells, with few minor variations in the initial steps. Despite all these advantages, this method uses different dangerous organic solvents and hence should be carefully handled. Moreover, this method is laborious and time-consuming compared to the other methods available, for example, column-based isolation of DNA.