GSM9061282: A549, RPSA_KO,  IAV_8h, DRIP-seq, rep 2 (Input); Homo sapiens; OTHER
Total nucleic acids were extracted from cells and tissues by SDS/proteinase K treatment followed by phenol_x0002_chloroform extraction and ethanol precipitation, some samples were extracted by DNA Kit. Typically more than 10 µg genomic DNA for each sample was fragmented by ultrasound (Perform 10-15 cycles of 30 s ON/ 30s OFF, spin after 3, 6, 9, and 12 cycles to ensure homogeneous sonication). The fragments length are limited within 500bp as possible. Followed by second DNA strand synthesis, dsDNA was cleaned up with 1.8 volume of DNA Beads and ligate “A” and adaptor. Add 10 µL ddH2O and 35 uL PEG/NaCl (1:1) to clean up and eluted with 10 µL ddH2O. Add 11 µL KAPA HiFi HotStart ReadyMix (2X) and 1 uL primers to amplify library. Each DNA library was amplified 9 -20 cycles depending on the starting material amount. Library was purified by 0.8 volume of DNA Beads. The library concentration and size distribution were checked by Qubit and 2100