GSM5692659: WT/TYLCCNV-IR-Bisulfite-seq_rep2; Nicotiana benthamiana; Bisulfite-Seq
Total DNA was extracted from infected plant leaves using Plant Genomic DNA Kit (TIANGEN, DP305).Total DNA (500 ng) was subjected to bisulfite treatment using EZ DNA Methylation Gold TM Kit (Zymo Research, D5048), according to the manufacturer's instructions. The bisulfite-treated DNA was used as templates for amplification of the IR regions of TYLCCNV or BSCTV. Primer sequences are listed in Supplementary Table 1. The products were purified with AMPure XP beads (Beckman, A63880) and the purified DNA products were PCR amplified and the adapters were added using GXL DNA polymerase (Takara, R050Q) and primer F (5'-AATGATACGGCGACCACCGAGATCTACAC-Index-TCGTCGGCAGCGTC-3') and primer R (5'-CAAGCAGAAGACGGCATACGAGAT-Index-GTCTCGTGGGCTCGG)96. The final products were purified with AMPure XP beads for sequencing on an Illumina HiSeqX-Ten platform by Annoroad Gene Technology (Beijing).