title: WGS of human Stool: Nm spikein 4
design: DNA was extracted from human stool using the Zymo quick-DNA fecal/soil DNA with 40-minute bead-bashing, and eluted into 1x TE buffer. An aliquot was set aside as a control. Internal controls (ICs) of A. hydrophila and L. pneumophila DNA (NIST RM 8376) were spiked-in at ~1e5 genome copy/uL and 1.4e4 genome copy/uL, respectively. A control aliquot was taken to check background signal and potential contamination from the IC addition. Separate dilution series were prepared for K. pneumoniae, N. meningitidis, E. faecalis, and L. monocytogenes DNA (RM 8376), starting at 1:10 dilution of the neat reference material into extracted stool DNA and making 6 total dilutions each (~1e6 copy/uL to ~10 copy/uL. NEBNext Ultra II FS was used to generate the libraries with unique dual indices plate.