GSM3401642: YJATAC0433: 5000 nuclei; Zea mays; ATAC-seq
Seeding maize tissue were chopped by a double edge razor blade and the slurry was filtered through Miracloth, followed by filtration through a 20 µm nylon mesh twice to remove large debris. After centrifugation, the nuclei were pelleted together with chloroplasts and starch grains. The pellet was resuspended in the CRB buffer where chloroplasts burst. Filter this nuclei suspension through a 10-µm mesh twice. Load approximately 400 ul of this green-colored nucleus suspension on top of the Percoll-Sucrose gradient and centrifuge the tube. Nuclei fractionated at the interface between CRB and SCB layer are transferred and pelleted in NEB buffer. Maize seedling genomic DNA was extract by MagMax Plant DNA Kit (Thermo Fisher Scientific). Freshly isolated nuclei and extracted gDNA were sujected to fragmentation and tagging. After cleanup, first PCR was performed to amplify DNA fragments and add barcodes. Second, the secondary PCR cycle numbers were determined by qPCR. The quality of constructed library was checked by Qubit and Bioanalyzer. Sequencing was performed with Illumina HiSeq 4000.
GEO Accession
GSM3401642