Illumina HiSeq 4000 paired end sequencing; Heart RNA-seq of control mice and mice with titin PEVK domain knockout, Rbm20 RNA recognition motif knockout, or double knockout
Male mice were kept on a 129S6 background and analyzed at 100 to 120 days of age. Hearts were rapidly excised after cervical dislocation without previous anesthesia, washed in PBS, and dissected. The tissue was snap frozen in liquid nitrogen and stored at -80C. Animals were kept at the Animal House of the Max-Delbrck Center for Molecular Medicine, Berlin. RNA from left ventricular tissue was isolated using TRIzol followed by a clean-up with the QIAGEN RNA micro kit.  cDNA libraries were generated using the Illumina TruSeq Stranded mRNA Sample Prep Kit.