Heart RNA-Seq of cardiomyocyte-specific and inducible Prmt5 knockout mouse model, nuclear sequencing without treatment
Heart RNA-Seq of cardiomyocyte-specific and inducible Prmt5 knockout mouse model, nuclear sequencing without treatment
Cardiomyocyte-specific nuclei isolation was performed via a fluorescence labeling of PCM1 and fluorescence-activated nuclei sorting. PCM1-positive nuclei were subjected to RNA extraction based on the RNeasy Micro Kit (Qiagen). Animals received Tamoxifen for KO induction (75mg/kg BW/d) followed by an Isoprenaline treatment (30mg/kg BW/d) PCM1-positive nuclei were subjected to RNA extraction based on the RNeasy Micro Kit (Qiagen). Library construction was performed according to the SmartSeq2 protocol (Picelli, S., Faridani, O., Björklund, Å. et al. Full-length RNA-seq from single cells using Smart-seq2. Nat Protoc9, 171–181 (2014). https://doi.org/10.1038/nprot.2014.006 and Bianca P Hennig, Lars Velten, Ines Racke, Chelsea Szu Tu, Matthias Thoms, Vladimir Rybin, Hüseyin Besir, Kim Remans, Lars M Steinmetz, Large-Scale Low-Cost NGS Library Preparation Using a Robust Tn5 Purification and Tagmentation Protocol, G3 Genes|Genomes|Genetics, Volume 8, Issue 1, 1 January 2018, Pages 79–89, https://doi.org/10.1534/g3.117.300257)