Purification of retrovirus genomic RNA suitable for chemical radioiodination.
An efficient method for the purification of genomic RNA from the retrovirus, caprine arthritis-encephalitis virus, is described. The method utilizes proteinase K, extraction with sodium perchlorate and chromatography on oligo(dT)-cellulose and results in highly purified RNA capable of being chemically iodinated with Na125 I to high specific radioactivity. The iodinated RNA exhibits 80-90% precipitability in 5% trichloroacetic acid and is greater than or equal to 99% sensitive to hydrolysis by ribonuclease. Several alternative methods which are effective for the preparation of eukaryotic ribosomal RNA are unreliable for purification of retrovirus RNA suitable for radioiodination.
D002845:Chromatography; D019286:Endopeptidase K; D010450:Endopeptidases; D005814:Genes, Viral; D007457:Iodine Radioisotopes; D008722:Methods; D010472:Perchlorates; D012367:RNA, Viral / Q000302:isolation & purification*; D012190:Retroviridae / Q000032:analysis*; D012260:Ribonucleases; D017670:Sodium Compounds
D007457:Iodine Radioisotopes; D010472:Perchlorates; D012367:RNA, Viral; D017670:Sodium Compounds; C031068:sodium perchlorate; D012260:Ribonucleases; D010450:Endopeptidases; D019286:Endopeptidase K