Carrier RNA enhancement of recovery of DNA from dilute solutions.
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 -20 degrees C 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 DNA's of heterogeneous molecular weight and conformation was also enhanced by the addition of carrier RNA. DNA's recovered by this method can be successfully digested with BamHI and ligated with T4 DNA ligase.
D011232:Chemical Precipitation; D004247:DNA / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000302:isolation & purification; D004270:DNA, Circular / Q000302:isolation & purification; D004278:DNA, Superhelical / Q000302:isolation & purification; D004279:DNA, Viral / Q000302:isolation & purification; D010957:Plasmids; D012313:RNA; D012331:RNA, Fungal; D012996:Solutions
D004269:DNA, Bacterial; D004270:DNA, Circular; D004278:DNA, Superhelical; D004279:DNA, Viral; D012331:RNA, Fungal; D012996:Solutions; D012313:RNA; D004247:DNA