PMC:4829102 / 37083-37954 JSONTXT

{"project":"2_test","denotations":[{"id":"26807646-24362840-73828213","span":{"begin":117,"end":119},"obj":"24362840"},{"id":"26807646-24362840-73828213","span":{"begin":117,"end":119},"obj":"24362840"},{"id":"26807646-24362840-73828214","span":{"begin":390,"end":392},"obj":"24362840"},{"id":"26807646-24362840-73828214","span":{"begin":390,"end":392},"obj":"24362840"},{"id":"26807646-24362840-73828215","span":{"begin":651,"end":653},"obj":"24362840"},{"id":"26807646-24362840-73828215","span":{"begin":651,"end":653},"obj":"24362840"},{"id":"26807646-24362840-73828216","span":{"begin":718,"end":720},"obj":"24362840"},{"id":"26807646-24362840-73828216","span":{"begin":718,"end":720},"obj":"24362840"}],"text":"ER-AsiSI resection assay\nThe level of resection adjacent to specific DSB (Chr 1: 89231183) was measured as described 40 with some modifications. Briefly ER-AsiSI U2OS cells were treated with 300 nM of 4-hydroxytamoxifen (4-OHT, Sigma) for 1 hour to allow the AsiSI enzyme to enter the nucleus and induce DSBs. Cells were then harvested and genomic DNA was extracted as previously described 40. Genomic DNA was then digested with the BsrGI enzyme or mock digested and was used as a template for qPCR performed using iTaq Universal Sybr Green Mix (Bio-Rad) and Rotor-Gene (Corbett Research) qPCR system. Primers used are listed in Supplementary Table 4 40. The percentage of ssDNA was calculated as previously described 40. All data were then related to the siControl treated sample, which was set as 100%. Statistical significance was determined with the Student’s t-test."}