title: GSM4284317: P2_ST_rep2; Homo sapiens; OTHER
library_construction_protocol: The Spatial Transcriptomics (ST) protocol was optimized for cSCC tissue  according to recommendations (Salmén et al., 2018). Once optimal conditions had been established, three cryosections per patient were cut at 10μm thickness onto spatial slides and processed immediately. For pre-permeabilization, sections were incubated at 37°C for 20 min with 0.5 U/µl collagenase (ThermoFisher) and 0.2 µg/µl BSA (NEB, Ipswich, MA) in HBSS buffer (ThermoFisher), washed with 0.1× SSC (Sigma-Aldrich), after which permeabilization was conducted at 37°C for 7 min in 0.1% pepsin (Sigma-Aldrich) dissolved in 0.1M HCl (Sigma-Aldrich). After incubation, the pepsin solution was removed and wells washed with 0.1× SSC. Reverse transcription and steps thereafter proceeded according to the ST protocol (Salmen et al., 2018). Second-strand cDNA synthesis was followed by in vitro transcription, adapter ligation, and a second RT. Sequencing handles and indexes were added in an indexing PCR and the finished libraries were purified and quantified as previously described (Ståhl et al., 2016)
attributes.tag: GEO Accession
attributes.value: GSM4284317