Fragmentation of DNA by Nebulization
This protocol was adapted from “Commonly Used Techniques in Molecular Cloning,” Appendix 8, in Molecular Cloning, Volume 3, 3rd edition (eds. Sambrook and Russell). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001.INTRODUCTION
DNA fragmentation is often necessary prior to library construction or subcloning for DNA sequencing. This protocol describes a method for DNA fragmentation by nebulization, in which the fine mist created by forcing a DNA solution through a small hole in the nebulizer unit is collected. The size of the fragments obtained by nebulization is determined chiefly by the speed at which the DNA solution passes through the hole, altering the pressure of the gas blowing through the nebulizer, the viscosity of the solution, and the temperature. Nebulization is easy, quick, and requires only small amounts of DNA (0.5-5 μg). The resulting DNA fragments are distributed over a narrow range of sizes (700-1330 bp). It requires ligation of DNA before nebulization and end-repair afterward.
