Title: Ultra-high throughput multiplexing and sequencing of >500 bp amplicon regions on the Illumina HiSeq 2500 platform

Johanna B. Holm; Michael S. Humphrys; Courtney K. Robinson; Matthew L. Settles; Sandra Ott; Li Fu; Hongqiu Yang; Pawel Gajer; Xin He; Elias McComb; Patti E. Gravitt; Khalil G. Ghanem; Rebecca M. Brotman; Jacques Ravel

doi:10.1101/417618

Title: Ultra-high throughput multiplexing and sequencing of >500 bp amplicon regions on the Illumina HiSeq 2500 platform

Johanna B. Holm, Michael S. Humphrys, Courtney K. Robinson, Matthew L. Settles, Sandra Ott, Li Fu, Hongqiu Yang, Pawel Gajer, Xin He, Elias McComb, Patti E. Gravitt, Khalil G. Ghanem, Rebecca M. Brotman, Jacques Ravel

School of Medicine

Research output: Contribution to journal › Article › peer-review

Abstract

Amplification, sequencing and analysis of the 16S rRNA gene affords characterization of microbial community composition. As this tool has become more popular and amplicon-sequencing applications have grown in the total number of samples, growth in sample multiplexing is becoming necessary while maintaining high sequence quality and sequencing depth. Here, modifications to the Illumina HiSeq 2500 platform are described which produce greater multiplexing capabilities and 300 bp paired-end reads of higher quality than produced by the current Illumina MiSeq platform. To improve the feasibility and flexibility of this method, a 2-Step PCR amplification protocol is also described that allows for targeting of different amplicon regions, thus improving amplification success from low bacterial bioburden samples.

Original language	English (US)
Journal	Unknown Journal
DOIs	https://doi.org/10.1101/417618
State	Published - Sep 14 2018

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Agricultural and Biological Sciences(all)
Immunology and Microbiology(all)
Neuroscience(all)
Pharmacology, Toxicology and Pharmaceutics(all)

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10.1101/417618

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Title : Ultra-high throughput multiplexing and sequencing of >500 bp amplicon regions on the Illumina HiSeq 2500 platform. / Holm, Johanna B.; Humphrys, Michael S.; Robinson, Courtney K.; Settles, Matthew L.; Ott, Sandra; Fu, Li; Yang, Hongqiu; Gajer, Pawel; He, Xin; McComb, Elias; Gravitt, Patti E.; Ghanem, Khalil G.; Brotman, Rebecca M.; Ravel, Jacques.

In: Unknown Journal, 14.09.2018.

Research output: Contribution to journal › Article › peer-review

Holm, Johanna B. ; Humphrys, Michael S. ; Robinson, Courtney K. ; Settles, Matthew L. ; Ott, Sandra ; Fu, Li ; Yang, Hongqiu ; Gajer, Pawel ; He, Xin ; McComb, Elias ; Gravitt, Patti E. ; Ghanem, Khalil G. ; Brotman, Rebecca M. ; Ravel, Jacques. / Title : Ultra-high throughput multiplexing and sequencing of >500 bp amplicon regions on the Illumina HiSeq 2500 platform. In: Unknown Journal. 2018.

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abstract = "Amplification, sequencing and analysis of the 16S rRNA gene affords characterization of microbial community composition. As this tool has become more popular and amplicon-sequencing applications have grown in the total number of samples, growth in sample multiplexing is becoming necessary while maintaining high sequence quality and sequencing depth. Here, modifications to the Illumina HiSeq 2500 platform are described which produce greater multiplexing capabilities and 300 bp paired-end reads of higher quality than produced by the current Illumina MiSeq platform. To improve the feasibility and flexibility of this method, a 2-Step PCR amplification protocol is also described that allows for targeting of different amplicon regions, thus improving amplification success from low bacterial bioburden samples.",

author = "Holm, {Johanna B.} and Humphrys, {Michael S.} and Robinson, {Courtney K.} and Settles, {Matthew L.} and Sandra Ott and Li Fu and Hongqiu Yang and Pawel Gajer and Xin He and Elias McComb and Gravitt, {Patti E.} and Ghanem, {Khalil G.} and Brotman, {Rebecca M.} and Jacques Ravel",

note = "Publisher Copyright: The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

year = "2018",

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doi = "10.1101/417618",

language = "English (US)",

journal = "Advances in Water Resources",

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AU - Holm, Johanna B.

AU - Humphrys, Michael S.

AU - Robinson, Courtney K.

AU - Settles, Matthew L.

AU - Ott, Sandra

AU - Fu, Li

AU - Yang, Hongqiu

AU - Gajer, Pawel

AU - He, Xin

AU - McComb, Elias

AU - Gravitt, Patti E.

AU - Ghanem, Khalil G.

AU - Brotman, Rebecca M.

AU - Ravel, Jacques

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PY - 2018/9/14

Y1 - 2018/9/14

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