Sequencing data of DNA repair substrate plasmid by TMEJ

<b>TMEJ assay in nuclear extracts</b>.The assay was performed as described by Dutta et al. (Dutta et al., 2017) with modifications. Briefly, exponentially growing U2OS cells transiently expressing POLQ-Flag-HA in 60 mm plates (90% confluent) were irradiated with X-rays (10Gy). After indicated time points of incubation, the irradiated and control cells were harvested for preparation of nuclear extracts. 100ng I-<i>Sce</i>I digested pBabehygro-EGFP-MMEJ (Truong et al., 2013), the repair substrate, mixed with 100μl nuclear extracts 30 min with gentle shaking at 30^oC followed by incubation at 15 h at 16^oC. 10μl of mixture were used to XL10- gold ultracompetent E. coli (Agilent Technologies) following the manufacturer’s protocol. The colonies in each agar plate were counted and submitted for PCR + sequence analysis of the product using the PCR primers: 5′-ACGGGGTCATTAGTTCATAGCCCA-3′, and 5′-GGGATTTTGCCGATTTCGGCC-3′; and the sequencing primer: 5'-ATGGTGAGCAAGGGCGAGGAG-3' (Genewiz Inc.).