An improvement on labelled messenger RNA preparation for DNA-RNA hybridisation.
An improved method for the removal of DNA and phenol residues after phenol extraction in m-RNA preparation has been developed. This involves repeated freeze-thaws of the aqueous layer under centrifugal force. As a model assay, five minutes pulse-labelled lambda cI47 late m-RNA is prepared and hybridised to the r-strain lambda DNA. After the centrifugation treatment, non-specific hybridisation is reduced to one-tenth of the untreated sample and the background with respect to input radioactivity for hybridisation is reduced to 0.3%. This low background m-RNA preparation is important especially in dealing with minor class m-RNA.
D010582:Bacteriophage lambda / Q000235:genetics; D002498:Centrifugation; D004279:DNA, Viral / Q000302:isolation & purification; D005615:Freezing; D008722:Methods; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification
D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral