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PacBio - Amplicon/16S Sequencing

In this workflow, one or more DNA fragments are selectively amplified. This allows multiplexing protocols to process many samples at once, reducing cost per sample significantly compared to whole genome sequencing protocols. However, some information is lost due to amplification, e.g. Methylation data. Amplicons up to 15kb can be sequenced. In order to pool different amplicons, they have to be of similar size.

A particular application of amplicon sequencing is 16S sequencing for the detection of microbial species from natural populations (microbiome metagenomics). Due to the long read length, the 16S genes can be completely sequenced, whereas short read sequencing techniques only cover certain variable ranges. This results in a significantly higher resolution for the species determination.

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