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 0,75g of purified DNA (sonication), followed by end-repair, A-tailing and ligation of methylated adaptors prior to custom probes (1:8 diluted) hybridization (16h). Bisulfite conversion was then performed with EZ-DNA Methylation-Gold Kit (ZYMO, Irvine, CA). 1st PCR was performed with 9 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.