Enzymatic Methyl sequencing (EM-seq) of primed and non-primed Arabidopsis thaliana under cold-stress
Enzymatic Methyl sequencing (EM-seq) of primed and non-primed Arabidopsis thaliana under cold-stress
RNA-seq was performed for 25-days-old Arabidopsis thaliana plants grown in control conditions (22°C) (non-primed, C), plants primed with cold stress at 4°C for 3 days (Priming only, P), plants triggered with cold stress at 4°C for 3 days (Triggering only, T), and the primed and triggered plants (Priming and Triggering, PT). Leaf samples of the plants were collected. Plants were grown in 0.5x Murashige and Skoog (MS) media with 0.8% agar with 0.1% sucrose content at 22°C with a long day (16 h light / 8 h dark) for 9 days, then transferred to soil for stress treatment and until the sample collection. Genomic DNA (gDNA) was extracted from 0.1g of frozen plant leaf tissue using the Nucleon PhytoPure DNA extraction kit (Cytiva, USA) according to the protocol manual from the manufacturer. EM-seq libraries were constructed using NEBNext Enzymatic Methyl-seq (EM-seq™) kit for Illumina (New England BioLabs, USA) and NEBNext® Multiplex Oligos for Enzymatic Methyl-seq (Unique Dual Index Primer Pairs) (New England BioLabs, USA) following the manufacturer's recommendations.