Abstract: The disclosure provides for single amplification and double amplification methods for preparing nucleic acid samples for sequencing.

Abstract: Rapid methods, capable of being performed in a single reaction tube, are described herein for constructing libraries for high-throughput polynucleotide sequencing applications, such as next generation sequencing (NGS) applications. Oligonucleotide probes include chemically-active groups at their 5? or 3? ends, or both, to facilitate the cleavage of their 5? or 3? ends, or both, following their hybridization to the single-stranded ends of frayed template fragments. Cleavage of probe ends reveal single-stranded regions at the ends of the hybridized fragments. Adaptors, specific to these ends, are ligated to the hybridized probe/template fragments, and blunt end fragments are ligated to blunt ends of hybridized probe/template fragments, if present, to generate the adaptor-ligated fragments of the library.

Abstract: The disclosure provides for single amplification and double amplification methods for preparing nucleic acid samples for sequencing.