Amplicon libraries
We use PCR to amplify 16S and ITS regions and attach unique indexes (or barcodes) to each product. We then pool the products into a library. In the future we will also develop the ability to target other loci (e.g., CO1 in animals).
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Our Protocol:16S and/or ITS Library Prep Protocol
Sequencing
We are sequencing using Illumina instruments, which all use a common chemistry. They differ some in the nature of the flow cells. The following video gives an overview of the sequencing by synthesis that happens on the Illumina instrument and would apply to the sequencing of one or more of our sequencing libraries. We have an iSeq device at UW for small scale work, and use sequencers elsewhere (e.g., a NovaSeq instrument at the GSAF at the University of Texas) for production work.
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