Determination of human DNA replication origin position and efficiency reveals principles of initiation zone organisation. Determination of human DNA replication origin position and efficiency reveals principles of initiation zone organisation

We describe a modification of initiation site sequencing (ini-seq, Langley et al., NAR 2016) in which density gradient centrifugation separates ‘heavy’ replicated DNA containing BrdU from DNA containing only ‘light’ dTTP before sequencing. This approach allows us to assign a replication initiation efficiency score to each of the 23905 origins identified. We also performed Short Nascent Strand sequencing (SNS-seq), an established method to map replication origins, in the same cell line as in Ini-seq2 for comparison. Overall design: Massive sequecing of human replication origin isolated from EJ30 cell line by either Ini-seq2 or SNS-seq method