Abstract
This chapter provides an overview of the S 1 nuclease protection assay. The assay allows the precise identification of pertinent gene boundaries in ribonucleic acid (RNA) transcripts. These can include intron/exon junctions and the 5´ or 3´ ends of transcripts. The S 1 nuclease protection assay allows a specific comparison between the RNA and a labeled DNA probe, which is a more stringent and sensitive method than the Northern blot analysis method but is also more tedious and requires more knowledge of the structure of the gene being expressed and analyzed. The Si nuclease assay works by the endonuclease digestion of single-stranded DNA. Thus, if the 32 P-DNA does not base pair precisely to the hybridizing RNA, extra tails or loops not protected by RNA hybridization will be excised. The chapter illustrates the use of denaturing polyacrylamide gel to determine the size of nondigested hybridized pieces. Resolution of the undigested S 1 -protected species yields detailed information about the regions of sequence homology between the probe and the protecting messenger RNA species.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
