Co-precipitation of protein and polyester as a method to isolate high molecular weight DNA.
DNA isolation is often the limiting step in genetic analysis using PCR and automated fragment analysis due to low quality or purity of DNA, the need to determine and adjust DNA concentrations after isolation etc. Several protocols have been developed which are either safe and provide good quality DNA or hazardous and provide excellent quality DNA. In this brief communication I describe a new and rapid method of DNA isolation which employs the co-precipitation of protein and polyester, in the presence of acetone, to remove contaminating proteins from a lysed-tissue sample, thus leaving high quality pure DNA. The advantages of this method are increased safety over the phenol:chloroform and the chaotrophic salt methods and increased purity over the salting-out method. Since the concentrations of DNA isolated using this method are relatively consistent regardless of the amount of starting tissue (within limits), adjustments of the DNA concentrations before use as templates in PCR's are not necessary.
Dixson
Jamie D
JD
A. E. Wood Fish Hatchery, Texas Parks and Wildlife Department, 507 Staples Road, San Marcos, TX 78666, USA. jamie.dixson@tpwd.state.tx.us
D000096:Acetone / Q000737:chemistry; D000818:Animals; D001492:Bass; D002397:Catfishes; D011232:Chemical Precipitation; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D029941:Fish Proteins / Q000737:chemistry; D005821:Genetic Techniques / Q000592:standards; D008970:Molecular Weight; D011091:Polyesters / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D015203:Reproducibility of Results
D029941:Fish Proteins; D011091:Polyesters; D011506:Proteins; D000096:Acetone; D004247:DNA