DNA reassociation using oscillating phenol emulsions.
Reassociating double-stranded DNA from single-stranded components is necessary for many molecular genetics experiments. The choice of a DNA reassociation method is dictated by the complexity of the starting material. Reassociation of simple oligomers needs only slow cooling in an aqueous environment, whereas reannealing the many single-stranded DNAs of complex genomic mixtures requires both a phenol emulsion to accelerate DNA reassociation and dedicated equipment to maintain the emulsion. We present a method that is equally suitable for reassociating either simple or complex DNA mixtures. The Oscillating Phenol Emulsion Reassociation Technique (OsPERT) was primarily developed to prepare heteroduplex DNA from alkali-denatured high molecular weight human genomic DNA samples in which hundreds of thousands of fragments need to be reannealed, but the simplicity of the technique makes it practical for less demanding DNA reassociation applications.
Bruzel
Alan
A
Department of Pediatrics, University of Pennsylvania, Philadelphia, PA 19104, USA.
Cheung
Vivian G
VG
D004247:DNA / Q000737:chemistry*; D004655:Emulsions; D009692:Nucleic Acid Heteroduplexes; D019800:Phenol / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry
D004655:Emulsions; D009692:Nucleic Acid Heteroduplexes; D019800:Phenol; D012965:Sodium Chloride; D004247:DNA