GSM9139246: WGBS, control, rep1; Solanum lycopersicum; Bisulfite-Seq
Genomic DNA isolation: DNA isolation from tomato leaves was performed in two biological replicates from each treatment and control groups from the first sampling time (S1, 48h) according to the Macherey-Nagel Nucleospin® Plant II kit protocol. Concentration and purity of the extracted DNA was measured on NanoPhotometer® N50 (Implen, Munchen, Germany). Firstly, the genomic DNA spiked with lambda DNA were fragmented to 200-400 bp. The DNA fragments were bisulfite treated to generate single strand DNA (ssDNA). During the bisulfite conversion, unmethylated cytosine were converted into uracil, while methylated cytosine staid unchanged. Methylation sequencing adapters were ligated, followed by double strand DNA synthesis. The library was ready after size selection and PCR amplification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount required.