title: Metabarcoding of marine sediments with microplastics: glass control day 16 replicate 3
design: 0.5 g of marine sediment were sonicated in 5 ml sterile phosphate buffer saline (PBS) pH 7.0 using a 15-sec pulse 5 times with 45-sec ice breaks and incubated at 65C overnight with proteinase K during 2 h. Subsequently, DNA was extracted with the DNEASY Powersoil Pro kit (Qiagen) following the manufacturer's instructions. Lysozyme (US Biological) was added prior to DNA extraction to all samples as a pretreatment and incubated at 37C for 30 min. A volume of 40 l of nuclease free water was used to resuspend the extracted DNA. Amplicons were generated with the 515F (5-GTGCCAGCMGCCGCGGTAA-3) and 806R (5-GGACTACHVGGGTWTCTAAT-3) primers and sequenced using a n Illumina NovaSeq 6000 by Novogene Co.