GSM8509590: IPFTMA5, Post-Xenium Visium HD (VUILD49LA, TILD113LA, TILD111LA, VUHD049); Homo sapiens; OTHER
Slides underwent probe hybridization overnight and probe ligation for 1hr at 37℃ following manufacturer's protocols. Prior to CytAssist probe capture, the Visium HD slide was washed an d sample slides were stained in eosin briefly before placement on the instrument. The probe transfer occurred on the CytAssist instrument for 30min at 37℃. Probes were then extended on the Visium HD slide and eluted from said slide into a tube. Pre-amplification and SPRIselect were performed according to manufacturer's protocols. (10X Demonstrated Protocol CG000685) Probe-based library was constructed using Dual Index Plate TS Set A (10X Genomics 1000251). Sample index PCR cycle number was determined by the Cq value from qPCR (Applied Biosystems QuantStudio 5). A final cleanup step with SPRIselect reagent was followed by storage at -20°C until sequencing. Next generation sequencing was carried out on Illumina's NovaSeq X platform using a 10 billion read, 300 cycle flow cell. Minimum sequencing recommendations were met using the product of tissue coverage percentage estimations and constant maximum read figure 275 million read pairs. Optimal cluster density was achieved with a lane loading concentration of 235 pM. (10X Genomics Demonstrated Protocol CG000685)