CARRIER RNA ENHANCEMENT OF RECOVERY OF DNA FROM DILUTE SOLUTIONS英文文献.pdfVIP

Vol. 144, No. 1,1987 BIOCHEMICALAND BIOPHYSICALRESEARCH COMMUNICATIONS April14,1987 Pages 27]-276 CARRIER RNA ENHANCEMENT OF RECOVERY OF DNA FROM DILUTE SOLUTIONS Margaret L. Gallagher, William F. Burke, Jr., and Karen Orzech Departmentof Botany and Microbiology, Arizona State University, Tempe, AZ 85287 Received February 26, 1987 SUMMARY. A study of the use of carrier RNA to improve precipitation of DNA from dilute solutions was conducted to define the conditions which optimize DNA recovery. Replicate samples containing labeled pBR322 and increasing concentrations of commercially-available Torula yeast RNA were ethanol precipitated at -20C for 1 h in microfuge tubes obtained from various manufacturers. Nucleic acids were pelleted by centrifugation for either 5 or 30 min, dried and resuspended. Although recovery was not identical in each type of microfuge tube, in all cases the percent recovery increased when carrier was added. In most cases, extending centrifugation to 30 min did not significantly increase recovery. Recovery of unlabeled DNAs of heterogeneous molecular weight and conformation was also enhanced by the addition of carrier RNA. DNAs recovered by this method can be successfully digested with BamHI and ligated with T4 DNA ligase. ~ 1987 Academic P. . . . . I n c . The precipitation of nucleic acids with ethanol in the presence of sodium