18S_meso5_week6
Water collected for eDNA analyses (1000 L) was filtered through 0.2 m polyethylsulfone filters (Sartorius - Type 15407- 47 mm) in the laboratory at the Universit du Qubec  Montral. The filters were then stored in -80 C prior to DNA extractions. DNA extractions were conducted with the Qiagen DNeasy PowerWater kit and protocols (Qiagen, 2017). Sequence amplification were done using QuantStudio 3D by ThermoFisher Scientific PCR machine. A one-step PCR was performed using the UCP HiFidelity PCR Kit from Qiagen (ready-to-use 2x concentrated master mix). Primers were at 0.25 M final concentration and we added between 2-4 l of DNA template. The PCR program is summarized in Table A4. We put 5 l on agarose gel before PCR clean-up, normalization and pooling. The Eukaryotic 18S gene was sequenced using the 960Fc (3-YRYRGGCTTAATTTGACTCAACRCG-5)-NSR1438 (5-GGGCATCACAG ACCTGTTAT-3) primer pair (Capo et al. 2016), with an Illumina Miseq analyzer of the CERMO platform at Universit du Qubec  Montral (UQAM).
SUB9704179
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