title: GSM5058585: Ischium; Homo sapiens; ATAC-seq
library_construction_protocol: Upon arrival the sample was dissected under a light dissection microscope. Pelvis elements were complete but then manually separated from one another for the ATAC protocol. Element samples were then sent out for sequencing running a pooled library for 3 rounds of sequencing on the Illumina NextSeq 500 in order to obtain approximately 50 million reads per sample. Samples were subjected to PCR amplification and barcoding following Buenrostro et al., 2015. 10 ul of transposed DNA was then placed in a reaction containing NEBNext High-Fidelity PCR Master Mix, ddH20, and primers. After amplification, samples were transferred to micro-centrifuge tubes and subjected to the OMEGA Bead Purification Protocol following manufacturers protocol. The samples were eluted in 30 ul of TE, run on a nanodrop, diluted to 5 ng/ul and run on a bioanalyzer. Prior to sequencing sample concentrations were determined using the KAPA Library Quantification Complete Kit (KK4824). Samples were then sent out to the Harvard University Bauer Core Facility for sequencing on one lane of the Illumina NextSeq 500, for three rounds, after which raw reads for each sample were pooled.