MOAseq of Zea mays: earshoot 0-1 cm , br2tr1
Nuclei were isolated from formaldehyde-fixed field-grown earshoots of Zea mays at the indicated developmental stage. Micrococcal Nuclease (MNase) was titrated on isolated nuclei and light digest conditions were selected based on the appearance of "nucleosomal ladders" on agarose gel electrophoresis following overnight decrosslinking, DNA purification by sequential extraction with Tris-saturated phenol, 1:1 Tris-saturated phenol:Chloroform, Chloroform, and finally precipitation with ethanol, sodium acetate, and linear polyacrylamide. Selected MNase digests were used as input for NGS library construction following manufacturer instructions (NEBNext Ultra II kit). Libraries were size-selected on Pippin HT 2% agarose with target range = 135-250 bp. Size-selected, uniquely indexed libraries were pooled in equimolar quantities and sequenced at the Florida State University College of Medicine on an Illumina HiSeq 2500.