SAM24440433
All steps were done according to 10x Protocol CG000495 Rev E. Temperature-regulated incubation steps were performed using a Bio-Rad C1000 Touch Thermal Cycler (Bio-Rad, Hercules, CA). Destained slides were immediately loaded into CytAssist Cassettes (11 mm gasket) for processing. Probe hybridization was performed for 16 hours using 10X Visium Mouse WT Probes (10x #2000455/2000456). Following hybridization, probe ligation and elongation steps were performed. A simple eosin stain was performed to enable visualization of the tissue, and the stained slides were loaded two at a time into the CytAssist instrument. CytAssist-enabled probe release deposited the probes onto CytAssist slides with two 11 mm capture areas. The CytAssist run conditions for all slides was 30 minutes at 37C. After a probe extension step, the probes were eluted from the CytAssist slides and collected into PCR tubes. A pre-amplification reaction was followed by SPRIselect-mediated cleanup (Beckman Coulter #B23319). Sample index PCR was performed using 10x Dual Index Plate TS Set A (10x #1000251) with 13 cycles. A final SPRI cleanup yielded a library for each capture area.