GSM8799789: VEH/SIV_3; Macaca mulatta; Bisulfite-Seq
Colon epithelial total DNA was extracted using DNeasy Blood and Tissue Kit (Qiagen Inc, CA). ~1 ug of DNA was used for reduced restriction bisulfite sequencing. Methyl-MiniSeq Library preparation was performed by Zymo Research. Libraries were prepared from 200-500ng of genomic DNA digested with 60 units of Taql and 30 units of MspI (NEB) sequentially and then extracted with DNA Clean & ConcentratorTM-5 (Zymo Research). Fragments were ligated to pre-annealed adapters containing 5'-methyl-cytosine according to Illumina's specified guidelines (www.illumina.com). Adapter-ligated fragments of 150-250bp and 250-300bp were recovered from a 2/5% NuSieve 1:1 agarose gel (Zymoclean Gel DNA Recovery Kit, Zymo Research). The fragments were then bisulfite treated using EX DNA Methylation-Lightning Kit (Zymo Research). Preparative-scale PCR was performed, and the resulting products purified for sequencing on an Illumina HiSeq.