Lakkasuo_23S_OLD3
DNA was extracted with the DNeasy PowerSoil Pro Kit (Qiagen). The lysis step was conducted on a TissueLyser II (Qiagen). Directly after elution, DNA was quantified on a Nanodrop ND-1000 spectrophotometer and subjected to sequencing. We used the primer pair P23SrV_f1: 5-GGA CAG AAA GAC CCT ATG AA-3 / P23SrV_r1: 5-CAG CCT GTT ATC CCT AGA G-3 (410 pb; Sherwood & Presting, 2007), targeting the 23S rRNA gene. The primers were tagged with Illumina adapters and used for DNA amplification. PCR was conducted in a total volume of 50 L, consisting of 25 L of AmpliTaq GoldTM Master Mix (Applied Biosystem, ThermoFisher), 19 L of nuclease-free ultrapure water, 1 L of forward and 1 L of reverse primer (20 M in final concentration), and 4 L of DNA. The following PCR conditions were used: 10 min at 95C (activation), followed by 35 cycles of 60 s at 94C (denaturation), 45 s at 55C (annealing), and 45 s at 72C (elongation), terminated by a final elongation of 10 min at 72C. The quality of PCR amplicons was assessed on a 1.65% agarose gel electrophoresis, and sequencing was performed using Illumina MiSeq technology (V3 chemistry, 2 x 250 pb) by the GeT-PlaGe platform (Genotoul Toulouse, France).