16S rRNAs (V4), replicate 2, chemocline layer (11m) in Cueva de la Mora lake
PCR reactions were set up with 11.375 uL sterile water, 10.625 uL of ex taq master mix with 806R reverse primer (0.2 M final concentration), 2 uL of previously extracted DNA (serial dilutions also used for troublesome samples), and 1 uL of 515F forward barcoded primer (0.2 M final concentration). Amplification with thermal cycler conditions: 3-minute denaturation at 94oC; then 35 cycles at 94oC for 45 s, annealing at 50oC (16S rRNA) and 57 oC (18S rRNA) for 60 s, and extension at 72oC for 90 s; followed by a 10-minute hold at 72oC for final extension of amplicons.