title: GSM7783008: Hep_Org_P0_4d [ATAC-seq]; Mus musculus; ATAC-seq
library_construction_protocol: Preparation of single hepatocyte organoid: Pipette up and down to destroy the cell/extracellular matrix mixture with a micropipette and then transfer the organoid suspension to a 15 mL centrifuge tube on ice. Optionally, turn the tube up and down from time to thaw the mixture thoroughly. Centrifuge for 5 min at 500-800 x g and 4 °C and discard the supernatant. Add 1 mL of trypsin solution to the pellet and pipette it to mix. Incubate at 37 °C for 5-10 min and then stop digestion by adding 2x the volume of wash medium. Centrifuge for 5 min at 800 x g and 4 °C and discard the supernatant. Resuspend the pellet with a culture medium and count the cells using a hemacytometer. ATAC-seq was performed as previously reported. Briefly, nucleus was extracted from samples, and the nucleus pellet was resuspended in the Tn5 transposase reaction mix. The transposition reaction was incubated at 37℃ for 30 min. Equimolar Adapter1 and Adatper 2 were added after transposition, PCR was then performed to amplify the library. After the PCR reaction, libraries were purified with the AMPure beads and library quality was assessed with Qubit.