Abstract
Hybridization capture is considered very cost- and time-effective method for enriching a massive amount of target loci distributed separately in a whole genome. However, divergent loci are difficult to enrich for the sequence mismatch between probes and target DNA. After analysis the distributional pattern of divergent loci in mitochondrial genomes (mitogenomes), we notice that the relatively variable regions are intercept by the relatively conservative regions. We propose to extend the length of library to overcome the problem. By using a home-made probe set to bait amphibian mitogeneomes DNA, we demonstrate that using 2 kb DNA libraries generate high sequence coverage in the highly variable regions than using 400 bp DNA libraries. These suggest that longer fragments in the library generally contain both relatively variable regions and relatively conservative regions. The divergent part DNA along with conservative part DNA is captured during hybridization. We present a protocol that allows users to overcome the gap problem for highly divergent gene capture.