![]() Alternatively, the target sequence itself can prime an RCR after its non-base paired 3' end has been removed by exonucleolytic activity. ![]() The displaced probe can then slip off the targetstrand and a rolling circle amplification is initiated. If the target strand has a nearby free 3' end, then the probe-target hybrids can be displaced by the polymerase used for replication. ![]() We demonstrate here that in order for the reaction to proceed efficiently, the probes must be released from the topological link that forms with target molecules upon hybridization and ligation. ![]() By using a rolling circle replication (RCR) mechanism, circularized but not unreacted probes can yield a powerful signal amplification. Circularizing oligonucleotide probes (padlock probes) have the potential to detect sets of gene sequences with high specificity and excellent selectivity for sequence variants, but sensitivity of detection has been limiting.
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