The Ion Torrent 16S Metagenomics Kit (Thermo Fisher Scientific, Pleasanton, CA, USA) was used to amplify 16S rRNA genes in all samples. Polymerase chain reaction (PCR) was performed with 2 primers according to the amplification conditions indicated in the manufacturer’s protocol. Primer 1 contained targets for the V2 (250 bp), V4 (288 bp), and V8 (295 bp) regions, whereas primer 2 contained the V3 (215 bp), V6–7 (260 bp), and V9 (209 bp) regions. After PCR amplification, the conjugated PCR products were purified according to the manufacturer’s instructions. DNA using the Ion Plus Fragment Library Kit (Thermo Fisher Scientific, Carlsbad, CA, USA) and Ion Xpress barcodes. The Ion Xpress Barcode Adapters 1-16 Kit (Thermo Fisher Scientific, Carlsbad, CA, USA) according to the manufacturer’s protocol was performed and the DNA library was purified. Each library was diluted to a concentration of 50 pM and pooled into equal volumes. The Ion 530 kit (Thermo Fisher Scientific) was used for the Ion Chef (Thermo Fisher Scientific) and Ion S5 XL platform (Thermo Fisher Scientific) workflows.
Ion torrent 16s metagenomics kit
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom
The Ion Torrent 16S Metagenomics Kit is a product designed for the identification and analysis of microbial communities in various samples. It provides a comprehensive solution for amplifying and sequencing the 16S rRNA gene, a widely used marker for microbial taxonomic classification.
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Lab products found in correlation
Ion plus fragment library kit, by Thermo Fisher Scientific (2 mentions)
Ion xpress barcode adapters 1 16 kit, by Thermo Fisher Scientific (1 mentions)
Ion chef, by Thermo Fisher Scientific (1 mentions)
Ionxpress barcodes, by Thermo Fisher Scientific (1 mentions)
Ion s5 xl platform, by Thermo Fisher Scientific (1 mentions)
Qubit dsdna hs assay kit, by Thermo Fisher Scientific (1 mentions)
Ion onetouch 2, by Thermo Fisher Scientific (1 mentions)
Ion s5, by Thermo Fisher Scientific (1 mentions)
Ion xpress barcode adapter, by Thermo Fisher Scientific (1 mentions)
Ampure xp bead, by Beckman Coulter (1 mentions)
2 protocols using ion torrent 16s metagenomics kit
1
16S rRNA Metagenomics Sequencing Protocol
2
Bacterial Identification Using 16S rRNA
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Bacterial identification was performed by sequencing the hypervariable regions of the 16S rRNA gene using two sets of primers that selectively amplify regions V2–4–8 and V3–6 and 7–9.
The 16S rRNA gene was amplified using an Ion Torrent 16S Metagenomics kit (Thermo Fisher Scientific Inc., Warrington, UK). The PCR products were tested by 2% agarose gel electrophoresis, purified with AMPure® XP Beads (Beckman Coulter Inc., Atlanta, GA, USA), and quantified using a QubitTM dsDNA HS Assay Kit (Invitrogen, Thermo Fisher Scientific Inc., Warrington, UK).
Libraries were then prepared with an Ion Plus Fragment Library Kit (Ion Torrent, Thermo Fisher Scientific Inc., Warrington, UK), indexing each sample with Ion Xpress ™ Barcode Adapters (Ion Torrent, Thermo Fisher Scientific Inc., Warrington, UK) according to the manufacturer’s protocol. The library pool was then clonally amplified by emulsion PCR using ion sphere particles (ISPs). Template preparation was performed on the Ion OneTouch™ 2 (Ion Torrent, Thermo Fisher Scientific Inc., Warrington, UK). Finally, sequencing was performed using a 530 chip on an Ion S5™ (Ion Torrent, Thermo Fisher Scientific Inc., Warrington, UK).
The 16S rRNA gene was amplified using an Ion Torrent 16S Metagenomics kit (Thermo Fisher Scientific Inc., Warrington, UK). The PCR products were tested by 2% agarose gel electrophoresis, purified with AMPure® XP Beads (Beckman Coulter Inc., Atlanta, GA, USA), and quantified using a QubitTM dsDNA HS Assay Kit (Invitrogen, Thermo Fisher Scientific Inc., Warrington, UK).
Libraries were then prepared with an Ion Plus Fragment Library Kit (Ion Torrent, Thermo Fisher Scientific Inc., Warrington, UK), indexing each sample with Ion Xpress ™ Barcode Adapters (Ion Torrent, Thermo Fisher Scientific Inc., Warrington, UK) according to the manufacturer’s protocol. The library pool was then clonally amplified by emulsion PCR using ion sphere particles (ISPs). Template preparation was performed on the Ion OneTouch™ 2 (Ion Torrent, Thermo Fisher Scientific Inc., Warrington, UK). Finally, sequencing was performed using a 530 chip on an Ion S5™ (Ion Torrent, Thermo Fisher Scientific Inc., Warrington, UK).
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