title: Illumina HiSeq 2500 paired end sequencing; Identification of Angustifolia 3 (ZmAN3) target genes through GSyellow-based ChIP-seq in Zea mays leaves
design: Identification of Angustifolia 3 (ZmAN3) target genes through GSyellow-based ChIP-seq in Zea mays leaves
library_construction_protocol: Leaf samples were always taken during steady state growth of the leaf (2d after the appearance of leaf4). Samples of four centimeter were pooled, encompassing the division zone, transition zone and the expansion zone. For each maize sample, 1.2 g leaf tissue (leaf 4 cm1-4) was fixed in Buffer A (Morohashi et al., 2012) for 15 min under vacuum conditions. For quenching, glycine was added to a final concentration of 0.1 M and incubated for 10 min. Fixed leaf tissue was washed with distilled water and frozen in liquid N2. ChIP-seq on PPD2-GSyellow and ZmAN3-GSyellow was performed using 5 L ab290 anti-GFP antibody (Abcam), as described in Morohashi et al., 2012. ChIP-seq libraries were prepared by GATC Biotech
attributes.tag: Experimental Factor: immunoprecipitate
attributes.value: AN3-Gsyellow tag