WGS of Trigonopterus: adult
Sequencing libraries were prepared with the Nextera DNA Flex  Library Prep kit (Illumina, San Diego, CA, USA). Procedures were followed the manufacturers protocol except that only half of the recommended volumes were used, i.e. starting with 15 l of genomic DNA template containing 1.5-5.0 ng DNA. Resulting libraries were quantified using a Qubit  3.0 Fluorometer with the dsDNA HS assay kit (Thermo Fisher Scientific, Waltham, MA, USA). Fragment distribution of libraries was examined using a Fragment Analyzer  dsDNA 910 kit (Agilent Technologies, Santa Clara, CA, USA) for a range of 35 to 1,500 bp. Based on the concentration and the average size distribution, the molar concentration was calculated for each sample. The twelve samples of this project were pooled in equimolar amounts together with other 84 samples and submitted to Macrogen Inc. (Seoul, South Korea) for sequencing on one lane of an Illumina Hiseq Xten for 2 x 150 bp. The resulting paired end fastq files were trimmed and quality filtered with BBduk: -Xmx1g in1=MZB0023_Nextera_1.fastq.gz in2=MZB023_Nextera_2.fastq.gz out=MZB0023_clean_Nextera.fastq ref=/home/alex/bbmap/resources/nextera.fa.gz ktrim=r k=23 mink=4 hdist=1 tpe tbo trimbyoverlap=t minoverlap=24 ordered=t qtrim=rl trimq=8