VF_1_1
For each water sample, 25 mL was filtered through 0.22 m pore size polycarbonate (PC) membrane filters (Millipore, Burlington, MA, USA). DNA was then extracted from the filters using the DNeasy PowerWater Kit (Qiagen, Hilden, Germany), according to the manufacturers instructions. DNA concentration and purity were determined using a NanoDrop spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA), a Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA), and 1% agarose gel electrophoresis. All DNA samples were stored at 20 C until further analysis. For each sample, extracted DNA was fragmented using Covaris S220 (Covaris, Woburn, MA, USA) to obtain c.a. 500-bp DNA fragments for the preparation of sequencing libraries. These were made with KAPA Hyper Prep Kit (Roche, Basel, Switzerland) and KAPA UDI Adapters Kit 15 uM (Roche, Basel, Switzerland). The quality of resulting libraries was assessed using Agilent 2200 TapeStation System and High Sensitivity D1000 reagents (Agilent, Santa Clara, CA, USA) and Kapa Library Quantification kit (Roche, Basel, Switzerland). Libraries were finally sequenced using the Illumina NovaSeq 6000 platform in the Genomics Core Facility at the Centre of New Technologies, University of Warsaw. The sequencing was conducted with NovaSeq 6000 S2 Reagent Kit v1.5 (300 cycles) in paired-end mode (2x150) according to the manufacturers instructions (Illumina Inc., San Diego, CA, USA). The sequencing generated from 16.6 to 35 million read pairs per sample.
SUB16000893
F_MDA_6_1