GSM2176274: Day5_CPC_control_ATAC-seq-Rep2; Mus musculus; OTHER
Control and Isl1 knockdown D5 cardiac progenitors were trypsinized and washed in PBS. 100,000 cells were further processed for ATAC library preparation using Tn5 Transposase form Nextera DNA Sample Preparation Kit (Illumina). Cell pellet was resuspended in 50µl PBS annd mixed with 25µl TD-Buffer, 2.5µl Tn5, 0.5µl 10% NP-40 and 22µl water. Cell/Tn5 mixture was incubeated at 37°C for 30min with occasional snap mixing. Transposase treatment was followed by 30min incubation at 50°C together with 500mM EDTA pH8.0 for optimal recovery of digested DNA fragments. For neutralisation of  EDTA 100µl of 50mM MgCl2 was added followed by purification of the DNA fragments by MinElute PCR Purification Kit (Qiagen) Tn5 Transposase form Nextera DNA Sample Preparation Kit (Illumina)