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16s metagenomic library preparation protocol

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The 16S metagenomic library preparation protocol is a laboratory procedure designed to generate sequencing-ready libraries from microbial DNA samples. This protocol enables the amplification and tagging of the 16S ribosomal RNA gene, which is commonly used to identify and characterize the composition of microbial communities.

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Lab products found in correlation

Miseq system, by Illumina (1 mentions) Miseq platform, by Illumina (1 mentions) V3 600 cycles kit, by Illumina (1 mentions)

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16S Metagenomics Sequencing Library Preparation protocol (13 citations) 16S library preparation protocol (4 citations) 16S metagenomics library preparation protocol (2 citations) 16S rRNA metagenomics sequencing library preparation protocol (2 citations)

2 protocols using 16s metagenomic library preparation protocol

1

16S Metagenomics Library Preparation

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Library preparation was performed by the JHMI Deep Sequencing and Microarray Core at the Johns Hopkins Medical Institute according to the Illumina 16S metagenomic library preparation protocol (Illumina, San Diego, CA) with primers that amplified the V4 region of the 16S gene. Paired-end sequencing of the pooled library was performed with the MiSeq system (Illumina, San Diego, CA), generating 2 × 250 reads. Adaptor and barcode trimming was also performed by the JHMI Deep Sequencing and Microarray Core at the Johns Hopkins Medical Institute.
DeLong K., Bensouda S., Zulfiqar F., Zierden H.C., Hoang T.M., Abraham A.G., Coleman J.S., Cone R.A., Gravitt P.E., Hendrix C.W., Fuchs E.J., Gaydos C.A., Weld E.D, & Ensign L.M. (2019). Conceptual Design of a Universal Donor Screening Approach for Vaginal Microbiota Transplant. Frontiers in Cellular and Infection Microbiology, 9, 306.
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2

Illumina MiSeq 16S rRNA Sequencing

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The purified amplified products were submitted to the ARC Biotechnology Platform, Onderstepoort, Pretoria, South Africa for library preparation and NGS on the Illumina MiSeq platform. Sequencing libraries were created according to the Illumina 16S metagenomic library preparation protocol available online (see Supplementary material). The 16S rRNA gene was sequenced on a MiSeq sequencer creating 300 bp paired-end reads with the V3 600 cycles kit (Illumina, San Diego, CA). Raw paired-end FASTQ files of each sample were received for subsequent analysis.
Geldenhuys J., Redelinghuys M.J., Lombaard H.A., Ehlers M.M., Cowan D, & Kock M.M. (2022). Diversity of the gut, vaginal and oral microbiome among pregnant women in South Africa with and without pre-eclampsia. Frontiers in Global Women's Health, 3, 810673.
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