title: eDNA of freshwater invertebrates: community
design: Nested-tagging for DNA metabarcoding: PCR1 in 25ul volume with 1x MyFi taq, 0.4uL of each Leray primers (CO1), 2ul of DNA. Clean-up 1 done as size-selection by magnetic beads following Quail et al. 2009. PCR2 following Illumina 16S protocol. Clean-up2 followed Quail et al 2009. Library loading conc was 15pM with 10% PhiX.