Rat Age
Rat Age
Rats were euthanised by a Schedule one procedure and the cardiac left ventricle dissected and snap frozen in RNALater. Naturally Ageing Aging Natural Ageing Aging RNA extraction was performed using the RNeasy Fibrous Tissues kit (Qiagen, UK) as per manufacturer's instructions. Approx. 30mg of tissue was lysed and homogenised using an ultra turrax (IKA Homogeniser, Sigma, UK) in buffer RLT before proteinase K digestion steps, total RNA bound followed by several washing steps and DNase digestion and total RNA eluted. Total RNA integrity was confirmed using the Bioanalyzer (Agilent Technologies, USA). Ribosomal RNA (rRNA) was depleted from RNA samples by using the Ribo-Zero™ rRNA Removal Kit (Epicentre, USA) in accordance with the instructions of the manufacturer. NEBNext® Ultra II Directional RNA Library Prep Kit for Illumina, so they are paired-end reads but the orientation of the RNA strand is preserved in the first strand