Pooled human saliva, unbound
The PowerSoil DNA isolation kit (MO BIO Laboratories, Carlsbad, CA) was used to extract DNA from 1ml of pooled human saliva (BioReclamation, Farmingdale, NY). The DNA was eluted in DpnI Binding Buffer and 400ng of the DNA was subjected to the DpnI pull-down assay.  The input, unbound, and bound/eluted fractions were used to prepare sequencing libraries. Library preparation and sequencing  The Nextera DNA Sample Preparation Kit (Illumina, San Diego, CA) was used to prepare libraries from input, unbound, and bound/eluted fractions from DpnI pulldown assays. Manufacturer’s instructions were followed for the library preparation except for recommended number of PCR cycles, which were varied according to the amount of DNA. Libraries were sequenced following the manufacturer’s instructions for the HiSeq 2500 Rapid Run mode to obtain 50 nucleotide readlengths.