GSM8945814: Ribo-seq from WT heart replicate 1; Mus musculus; OTHER
Cells or pulverized tissue samples were lysed in lysis buffer containing 100 μg/mL cycloheximide on ice. Lysates were clarified by centrifugation and treated with RNase I to generate ribosome-protected fragments (RPFs). RPFs were isolated using the Epi™ Ribosome Profiling Kit (Epibiotek, R1814), followed by purification using the RNA Clean & Concentrator-5 kit (ZYMO, R1016). rRNA was removed using the Epi™ RiboRNA Depletion Kit (Human/Mouse/Rat) (Epibiotek, R1805). Sequencing libraries were constructed from purified ribosome-protected fragments (RPFs) using the QIAseq miRNA Library Kit (QIAGEN, 1103679) according to the manufacturer's protocol. Final libraries were quantified, quality-checked, and subjected to high-throughput sequencing on the SURFSeq 5000 platform.