ROLLING CIRCLE AMPLIFICATION ASSAY FOR NUCLEIC ACID ANALYSIS

摘要

Method and reagents for analysis of nucleic acid sequences is disclosed. In this method a plurality of padlock probes is provided. Each padlock probe may hybridize to a locus on a target nucleic acid under hybridization conditions. If a targeted variant is present at the locus, the padlock probe may be ligated to form an amplification target circle. The amplification target circle acts as a template for production of tandem-sequence DNA. The tandem-sequence DNA may then be digested into non-tandem detection fragments which are subsequently separated and detected. The plurality of padlock probes are designed such that ligation of the probes, amplification of the target circle, and digestion of the tandem-sequence DNA subsequently produced, and detection may all be effected with the same set of reagents. Each probe targets a unique locus variant on the target nucleic acid sequence and produces a detection fragment that may be distinguished from detection fragments produced from other padlock probe in the plurality of padlock probes by using a fragment analysis detector.