Enrichment of megabase-sized DNA molecules for single-molecule optical mapping and next-generation sequencing.

ABSTRACT: Next-generation sequencing (NGS) has caused a revolution, yet left a gap: long-range genetic information from native, non-amplified DNA fragments is unavailable. It might be obtained by optical mapping of megabase-sized DNA molecules. Frequently only a specific genomic region is of interest, so here we introduce a method for selection and enrichment of megabase-sized DNA molecules intended for single-molecule optical mapping: DNA from a human cell line is digested by the NotI rare-cutting enzyme and size-selected by pulsed-field gel electrophoresis. For demonstration, more than 600 sub-megabase- to megabase-sized DNA molecules were recovered from the gel and analysed by denaturation-renaturation optical mapping. Size-selected molecules from the same gel were sequenced by NGS. The optically

SUBMITTER: Lopacinska-Jorgensen JM 

PROVIDER: S-EPMC5738345 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature