GSM2876491: OYH_RNAseq; Crassostrea gigas; RNA-Seq
Hepatopancreas was dissected manually from adult oysters. RNAseq:Total RNA was extracted following standard TRIzol protocol. Sequencing libraries were prepared using Illumina Nextera XT library preparation kit (FC-131-1024). ATACseq:Fresh hepatopancreas tissue fragments were incubated in 0.8 mg/ml collagenase P (Sigma, 11213857001) at room temperature for 20 min and gently pipetted every 5 min to release single cells. The cell suspension was filtered through a 40 μm cell strainer (BD Falcon, USA), transferred to a new tube, and centrifuged for 5 min at 1,000 rpm. The supernatant were discarded, and the pellet suspended in MDPBS for cell viability analysis and cell counting.  50,000 cells were collected at 3,000 rpm for 5 min, 4°C. The transposition reaction, purification, library construction and quantitation followed established methods, including tagmentation using a Nextera DNA kit (Illumina FC-121-1030 ) for 30 minutes at 37°C, amplification of tagmented DNA using NEB Next High-Fidelity 2X PCR Master Mix for 11 cycles and efficiency assessed using Agilent Tapestation. RNAseq: Standard TruSeq library construction protocol