Targeted genome bisulfite sequencing of Populus nigra : cambium of C001POK
Custom targeted genome bisulfite sequencing was performed using the SureSelect XT Methyl-Seq Target Enrichment (Agilent, Santa Clara, CA, USA,) according to Agilent recommendations. The workflow of the library preparation started with fragmentation to 200 base pairs (bp) of 3 g of purified DNA (sonication), followed by end-repair, A-tailing and ligation of methylated adaptors prior to custom probes (1:1 diluted) hybridization (16h). Bisulfite conversion was then performed with EZ-DNA Methylation-Gold Kit (ZYMO, Irvine, CA). 1st PCR was performed with 8 cycles, 2nd PCR (indexing) was performed with 6 cycles. Samples to be sequenced were combined at equimolar concentrations in a single pool and the sequencing was performed in paired ends (2 x 150 pb) on 1 lane of an Illumina NovaSeq6000 S4 flow-cell.