GSM5268645: P1ShamD7; Mus musculus; OTHER
Hearts at 3-day or 7-day post the surgery were dissected and immediately embedded in a tissue embedding mold filled with OCT and placed in a bath of isopentane and liquid nitrogen. OCT embedded heart tissue blocks were cryosectioned in a cryostat to generate appropriately sized sections for Visium Spatial slides. 10 um sections at 200 um below the ligation suture were collected and placed on the Visum Spatial slides capture areas. Heart sections were permeabilized for 18 min followed by RNA reverse transcription. The optimized permeabilization time was determined using the Visum Spatial Tissue Optimization Kit (10xGenomics) according to the manufacturer's protocol. Second strand cDNA was subsequently synthesized on tissue sections and was transferred to an Eppendorf tube after denaturation. The cDNA from the tissue slide that was spatially barcoded was next amplified via PCR and enzymatically fragmentated. P5, P7, i7 and i5 sample indexes are added via End Repair, A-tailing, Adaptor ligation and PCR to make the final sequencing library.