Locus-patterned sequence oriented enrichment for multi-dimensional gene analysis

Abstract

Multi-dimensional gene analysis provides in-depth insights into gene sequence, locus variations and molecular abundance, whereas it is vulnerable to the perturbation of complex reaction networks and always compromises on the discrimination of analogous sequences. Here, we present a sequence oriented enrichment method patterned by the prescribed locus without crosstalk between concurrent reactions. Energetically favourable structures of nucleic acid probes are theoretically derived and oriented to a specific gene locus. We designed a pair of universal probes for multiple conserved loci to avoid side reactions from undesired interactions among increased probe sets. Furthermore, competitive probes were customized to sink analogues for differentiating the reaction equilibrium and kinetics of sequence enrichment from the target, so variant loci can be synchronously identified with nucleotide-level resolution. Thus, the gene locus guides sequence enrichment and combinatorial signals to create unique codes, which provides access to multidimensional and precise information for gene decoding.