title: qPCR amplicon of Hemidactylium scutatum intergenic spacer region: Three Mile Bay, Sample 11
design: We modified Four-toed salamander qPCR primers by extending the forward and reverse qPCR primer sequences to include Illumina MiSeq adapter sequences, a unique barcode, and a primer pad sequence which were joined to the qPCR primer by a linker sequence. Amplicon samples were pooled, cleaned wtih QIAquick PCR purification kit (Qiagen), combined in an equimolar pools, spiked with 15% PhiX, and sequenced with custom primers on an Illumina MiSeq nano chip with v2 chemistry and 2x 150bp read lengths.