Truseq nano dna lt library

The genomic DNA of the phage was extracted using the Norgen Biotek Phage DNA Isolation Kit (Norgen Biotek Corp. Thorold, ON, Canada). Briefly, phage genomic DNA was obtained through 4 steps of propagating the phage in liquid cultures and preparing the phage supernatant containing 10⁹ PFU/mL, lysate preparation, sample binding to column, column washing, and DNA elution. Genomic DNA sequencing was performed using the Illumina Novaseq platform (Shanghai Personalbio Technology Co., Ltd. Shanghai, China). The standard Illumina TruSeq Nano DNA LT library preparations (Illumina TruSeq DNA Sample Preparation Guide) were used to construct the required online genome libraries. Genemarks [24 (link)] software was used to predict the protein-coding genes of the genome, and the sequence alignment of the coding genes was completed by Diamond software [25 (link)]. The putative protein function of the ORFs was annotated using the NR database. Diamond BLASTp was used to compare the protein sequences encoded by genes with the protein sequences in the database, to identify the function of the presumed proteins. Non-coding RNAs in the genome were predicted by comparing the ncRNA annotation database http://rfam.xfam.org/ (accessed on 13 August 2021) [26 (link)]. Phylogenetic analysis was conducted using MEGA (version X _10.2.6, Auckland, New Zealand).

Before DNA extraction, worms were treated with 7.5% (wt/vol) N-acetyl cysteine in PBS for 10 minutes, followed by PBS-only wash for 5 minutes. DNA was extracted using the Gentra Puregene Tissue Kit (Qiagen, #158667). A TruSeq Nano DNA LT library (Illumina, 125bp, paired-end) was constructed and sequenced at the UW-Madison Biotechnology center on a HiSeq 2500 platform.