GSM9380660: Tonsil, HPV-positive, sample1; Homo sapiens; OTHER
Two consecutive tissue sections of Normal tonsil tissue and HPV-positive tissue 10 μm were trimmed. One slide was stained by H&E staining. The second slide was affixed to the Stereo-seq chip surface and incubated at 37°C for 3-5 minutes. Subsequently, the sections were methanol-fixed and subjected to a 40-minute incubation at -20°C. The Stereo-seq library preparation and sequencing were conducted following a previously published protocol Qubit™ dsDNA HS Assay Kit (ThermoFisher, Q32854) quantified for the concentrations of the resulting PCR products. Then TME (Stereo-seq Library Preparation kit, STOmics) at 55 °C for 10 min fragmented a total of 20 ng of DNA, after which the reactions were then ceased with Stop Buffer (Stereo-seq Library Preparation kit, STOmics). Fragmented products were amplified with PCR Library Mix (Stereo-seq Library Preparation kit, STOmics) as follows1 cycle of incubation at 95 °C for 5 min, 13 cycles at 98 °C for 20 s, 58 °C for 20 s and 72 °C for 30 s, and a final incubation at 72 °C for 5 min. AMPure XP Beads (0.6× and 0.15×) (Stereo-seq Library Preparation kit, STOmics) was used to purify the PCR products. Qubit dsDNA HS Kit (ThermoFisher, Q32854) measured the concentration of purified PCR product. The DNB was prepared by High-throughput Sequencing Primer Kit (STOmics) and sequenced on MGI DNBSEQ-G400 sequencer.