mNGS: Target enrichment: Whole Blood: 003-D0
Cultured Newcastle disease virus (NDV), an RNA virus of a non-human origin, was added to the whole blood samples at concentrations ranging between 110^3 to 110^4 particles per 1 ml of whole blood as a positive internal control. QIAamp ccfDNA/RNA kit (Qiagen, Cat#55184) and AllPrep DNA/RNA Mini kit (Qiagen, Cat#80204) were used to extract total nucleic acid from plasma and white blood cells, respectively. SuperScript III kit (Invitrogen Life Technologies) was used for cDNA synthesis and Klenow fragment (3-5exonuclease negative) (New England Biolabs) was used for second-strand synthesis. The library was constructed according to SeqCap EZ HyperCap workflow users guide (Version 2.3, Roche). SeqCap EZ VirCapSeq-VERT Panel was used to perform target enrichment during library preparation.