GSM5618500: Flax variety JRF-2 seedlings treated with 5-Azacytidine - heat stress exposed; Linum usitatissimum; Bisulfite-Seq
Shoot apices (3-5 cm) of 10 randomly chosen treated and untreated seedlings were sample-wise pooled and extracted genomic DNA using the DNeasy Blood and Tissue Kit (Qiagen Inc., USA) as per the manufacturer's instructions 1µg of purified genomic DNA was treated with Bisulfite conversion kit from Active Motif Inc., as per the manufacturer's instructions. About 200ng of Bisulfite converted genomic DNA was size fractionated (450–500 bp) in sonicator. Ligation of sequencing adapters and paired-end (PE) sequencing libraries were prepared using the NEBNext® Ultra™ DNA Library Prep Kit for Illumina® (New England Biolabs Inc.) as per manufacturer's instructions. A high-fidelity PCR amplication  of the librraies was performed using NEBNext High-Fidelity PCR Master Mix (New England Biolabs Inc.) to ensure enrichment of libraries. The amplified libraries were quality checked on Bioanalyzer 2100 using the High Sensitivity (HS) DNA kit (Agilent Technologies Inc.) as per manufacturer's instructions. Cluster generation and paired-end library sequencing performed on Illumina HiSeq 2500 platform (2x150bp chemistry).