title: Small RNA Library Preparation Method for Next-Generation Sequencing Using Chemical Modifications to Prevent Adapter Dimer Formation.
abstract: For most sample types, the automation of RNA and DNA sample preparation workflows enables high throughput next-generation sequencing (NGS) library preparation. Greater adoption of small RNA (sRNA) sequencing has been hindered by high sample input requirements and inherent ligation side products formed during library preparation. These side products, known as adapter dimer, are very similar in size to the tagged library. Most sRNA library preparation strategies thus employ a gel purification step to isolate tagged library from adapter dimer contaminants. At very low sample inputs, adapter dimer side products dominate the reaction and limit the sensitivity of this technique. Here we address the need for improved specificity of sRNA library preparation workflows with a novel library preparation approach that uses modified adapters to suppress adapter dimer formation. This workflow allows for lower sample inputs and elimination of the gel purification step, which in turn allows for an automatable sRNA library preparation protocol.
authors.affiliation: Research and Development, Cell and Molecular Biology, TriLink BioTechnologies LLC., San Diego, California, United States of America.
authors.forename: Sabrina
authors.identifier: http://orcid.org/0000-0002-6848-1088
authors.initials: S
authors.lastname: Shore
authors.affiliation: Research and Development, Cell and Molecular Biology, TriLink BioTechnologies LLC., San Diego, California, United States of America.
authors.forename: Jordana M
authors.initials: JM
authors.lastname: Henderson
authors.affiliation: Research and Development, Cell and Molecular Biology, TriLink BioTechnologies LLC., San Diego, California, United States of America.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Lebedev
authors.affiliation: Research and Development, Cell and Molecular Biology, TriLink BioTechnologies LLC., San Diego, California, United States of America.
authors.forename: Michelle P
authors.initials: MP
authors.lastname: Salcedo
authors.affiliation: Research and Development, Cell and Molecular Biology, TriLink BioTechnologies LLC., San Diego, California, United States of America.
authors.forename: Gerald
authors.initials: G
authors.lastname: Zon
authors.affiliation: Research and Development, Cell and Molecular Biology, TriLink BioTechnologies LLC., San Diego, California, United States of America.
authors.forename: Anton P
authors.initials: AP
authors.lastname: McCaffrey
authors.affiliation: Engineering and Instrumentation, Synthetic Genomics, Inc., La Jolla, California, United States of America.
authors.forename: Natasha
authors.initials: N
authors.lastname: Paul
authors.affiliation: Research and Development, Cell and Molecular Biology, TriLink BioTechnologies LLC., San Diego, California, United States of America.
authors.forename: Richard I
authors.initials: RI
authors.lastname: Hogrefe
mesh_terms: D052157:Aptamers, Nucleotide; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing; D058727:RNA, Small Untranslated / Q000737:chemistry* / Q000235:genetics*
chemical_list: D052157:Aptamers, Nucleotide; D058727:RNA, Small Untranslated