title: Facile, high quality sequencing of bacterial genomes from small amounts of DNA.
abstract: Sequencing bacterial genomes has traditionally required large amounts of genomic DNA (~1 μg). There have been few studies to determine the effects of the input DNA amount or library preparation method on the quality of sequencing data. Several new commercially available library preparation methods enable shotgun sequencing from as little as 1 ng of input DNA. In this study, we evaluated the NEBNext Ultra library preparation reagents for sequencing bacterial genomes. We have evaluated the utility of NEBNext Ultra for resequencing and de novo assembly of four bacterial genomes and compared its performance with the TruSeq library preparation kit. The NEBNext Ultra reagents enable high quality resequencing and de novo assembly of a variety of bacterial genomes when using 100 ng of input genomic DNA. For the two most challenging genomes (Burkholderia spp.), which have the highest GC content and are the longest, we also show that the quality of both resequencing and de novo assembly is not decreased when only 10 ng of input genomic DNA is used.
authors.affiliation: Los Alamos National Laboratory, P.O. Box 1663, MS M888, Los Alamos, NM 87545, USA.
authors.forename: Momchilo
authors.initials: M
authors.lastname: Vuyisich
authors.affiliation: Los Alamos National Laboratory, P.O. Box 1663, MS M888, Los Alamos, NM 87545, USA.
authors.forename: Ayesha
authors.initials: A
authors.lastname: Arefin
authors.affiliation: Los Alamos National Laboratory, P.O. Box 1663, MS M888, Los Alamos, NM 87545, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Davenport
authors.affiliation: Los Alamos National Laboratory, P.O. Box 1663, MS M888, Los Alamos, NM 87545, USA.
authors.forename: Shihai
authors.initials: S
authors.lastname: Feng
authors.affiliation: Los Alamos National Laboratory, P.O. Box 1663, MS M888, Los Alamos, NM 87545, USA.
authors.forename: Cheryl
authors.initials: C
authors.lastname: Gleasner
authors.affiliation: Los Alamos National Laboratory, P.O. Box 1663, MS M888, Los Alamos, NM 87545, USA.
authors.forename: Kim
authors.initials: K
authors.lastname: McMurry
authors.affiliation: Los Alamos National Laboratory, P.O. Box 1663, MS M888, Los Alamos, NM 87545, USA.
authors.forename: Beverly
authors.initials: B
authors.lastname: Parson-Quintana
authors.affiliation: Parkview Medical Center School for Medical Laboratory Science, 400 West 16th Street, Pueblo, CO 81003, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Price
authors.affiliation: Los Alamos National Laboratory, P.O. Box 1663, MS M888, Los Alamos, NM 87545, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Scholz
authors.affiliation: Los Alamos National Laboratory, P.O. Box 1663, MS M888, Los Alamos, NM 87545, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Chain