Barcoding of Chytridiomycota sp.
Genomic DNA was extracted using the peqGOLD Tissue DNA Mini Kit. PCRs were conducted in 25- l reactions using 0.5  l Herculase II Fusion enzyme (Agilent Technologies), 5  l of 5x PCR buffer, 0.62  l each primer (10 uM), 0.25  l dNTPs (250 mM each), 0.3  l BSA (20mg/ml BSA) on a SensoQuest labcycler with 2 min denaturation at 95  C, 13-30 cycles of 94  C for 30 sec, 55  C for 30 sec and 70  C for 4 min, and a final elongation at 70  C for 10 min. Multiple PCR reactions (up to 50) were required for each environmental sample to ensure sufficient product for library preparation (1 g purified PCR product).