title: A cost-effective RNA sequencing protocol for large-scale gene expression studies.
abstract: RNA sequencing has increasingly become an indispensable tool for biological research. While sequencing costs have fallen dramatically in recent years, the current cost of RNA sequencing, nonetheless, remains a barrier to even more widespread adoption. Here, we present a simple RNA sequencing protocol with substantially reduced costs. This protocol uses as little as 10 ng of total RNA, allows multiplex sequencing of up to 96 samples per lane, and is strand specific. Extensive validation using human embryonic stem cells showed high consistency between technical replicates at various multiplexing levels.
authors.affiliation: Morgridge Institute for Research, Madison, Wisconsin, United States of America.
authors.forename: Zhonggang
authors.initials: Z
authors.lastname: Hou
authors.affiliation: Morgridge Institute for Research, Madison, Wisconsin, United States of America.
authors.forename: Peng
authors.initials: P
authors.lastname: Jiang
authors.affiliation: Morgridge Institute for Research, Madison, Wisconsin, United States of America.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Swanson
authors.affiliation: Morgridge Institute for Research, Madison, Wisconsin, United States of America.
authors.forename: Angela L
authors.initials: AL
authors.lastname: Elwell
authors.affiliation: Morgridge Institute for Research, Madison, Wisconsin, United States of America.
authors.forename: Bao Kim S
authors.initials: BK
authors.lastname: Nguyen
authors.affiliation: Morgridge Institute for Research, Madison, Wisconsin, United States of America.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Bolin
authors.affiliation: Morgridge Institute for Research, Madison, Wisconsin, United States of America.
authors.forename: Ron
authors.initials: R
authors.lastname: Stewart
authors.affiliation: 1] Morgridge Institute for Research, Madison, Wisconsin, United States of America [2] Department of Cell and Regenerative Biology, University of Wisconsin-Madison, Madison, Wisconsin, United States of America [3] Department of Molecular, Cellular, and Developmental Biology, University of California Santa Barbara, Santa Barbara, California, United States of America.
authors.forename: James A
authors.initials: JA
authors.lastname: Thomson
mesh_terms: D003362:Cost-Benefit Analysis; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000191:economics / Q000379:methods*; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000191:economics / Q000379:methods*; D006801:Humans; D015203:Reproducibility of Results; D017423:Sequence Analysis, RNA / Q000191:economics / Q000379:methods*
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