GSM8816653: KOIR; Mus musculus; OTHER
Fresh frozen tissue were OCT embedded, and 10um frozen sections were cut with cryostat Reverse transcription (RT), second-strand cDNA synthesis, adaptor ligation and a second RT was generated and libraries were constructed according to the performer's protocol. Sequencing handles and indexes were added in an indexing PCR and the finished libraries were purified and quantified. Sequencing was performed on the Illumina NovaSeq 6000 with a sequencing depth of at least 50,000 reads per sopt(100μm) and 150bp (PE150) paired-end reads (performed by Biomarker Technologies Corporation, Beijing, China).Primer spots were stained by hybridization of fluorescently labeled probes and imaged on the Metafer Slide Scanning platform. The resulting spot image was loaded into the BSTMatrix and BSTViewer along with the previously obtained BF tissue image of the same area. The two images were aligned and the built-in tissue recognition tool was used to extract spots covered by tissue.