Amplicon library preparation
Guidelines for amplicon library preparation for high-throughput sequencing on the Illumina platform. The documentation provides marker-specific protocols for 16S (bacteria), ITS2 (fungi), and COI (arthropods). Each protocol uses a 2-step PCR design: a first PCR amplifies the target region with primers carrying Illumina sequencing adaptor sites and shifters; a second PCR adds P5/P7 indexing for multiplexing. Consumables, thermocycling conditions, and cleanup steps are described in detail for each marker; the overall workflow is shared across markers, with differences mainly in primer pairs and 1st PCR annealing temperatures.
16S
ITS2
COI
18S
12S
