GSM7663226: END-seq, CPT 10 µM, Rep1; Homo sapiens; OTHER
Cells were embedded in agarose plugs and, after proteinase K and RNase A digestion, DSBs are recovered by ligation of a biotinylated adapter. DNA is then extracted from plug and sheared by sonication. Sheared fragments containing the biotinylated adapter are captured with streptavidin-coated beads. A second adapter is ligated to the other end of the fragments and library amplified with Illumina barcoded p5 and p7. End-Seq libraries were prepared incorporating Illumina barcoded p5 and p7 with KAPA HiFi HotStart Ready Mix by PCR. All the recovered DNA fragments were used for amplification. Libraries were amplified using 16 cycles on the thermocycler.