title: C24_x_Seattle_R2_S25
design: Genomic DNA was extracted from a single flash frozen rosette leaf using DNeasy Plant Mini Kit (Qiagen, Switzerland). Between 200-300 ng of genomic DNA was bisulfite-converted in a 130-microliter conversion reagent in a thermal cycler (98 C for 15 min, 65 C for 3 h, and 4 C, up to 20 h or overnight) and subsequently cleaned with the EZ DNA Methylation-Gold Kit (Orange, CA) following the manufacturers instructions. Next, bisulfite-converted ssDNA was used to make Illumina libraries with commercial xGenTM Methylation-Sequencing DNA Library Preparation Kit (IDT, Leuven, Belgium) following the manufacturer's instructions and indexed with combinatorial dual indices from the same company. Libraries were amplified with 9 PCR cycles and eluted in 20 microliter of elution buffer, quantified and validated using High Sensitivity D1000 Screen Tape (Agilent Technology, Basel, Switzerland). All the pre-made multiplexed libraries were sequenced on an Illumina NovaSeq X Plus instrument using 2 x 150 PE reads (Illumina, USA).
library_name: C24_x_Seattle_R2_S25