Illumina HiSeq 4000 paired end sequencing; Epigenetic atlas of ageing mouse heart
Epigenetic atlas of ageing mouse heart
Mice were obtained from Janvier (Le Genest Saint-Isle, France) and all animal experiments were approved by the Regional Board of the State of Hessen, Germany. The obtained hearts were either snap frozen with liquid nitrogen for nuclei isolation or freshly used for cell isolation.  Cross-sections of mouse hearts (derived from three 12 weeks old and from three 18 months old mice) were dissociated in liquid nitrogen using a mortar. The sample was then transferred into a Dounce homogenizer. For isolation of nuclei we strictly followed the experimental steps that are detailed in 3. Samples were homogenized and lysed to release the nuclei, followed by single-cell FACS (Aria III, BD Genomics) sorting of DAPI stained high quality nuclei to sort out any residual cell debris. Integrity and purity of the nuclei was inspected visually using a Leica DMi8 inverted microscope. Yield of nuclei was quantified by using a Neubauer chamber.  Following Smart-seq2 protocol