GSM9693387: Peyer's patch immune cells, pooled 16 mice, ScaleBio scRNA-seq multiplexed library_1; Mus musculus; RNA-Seq
Trif- and Il1r1-deficient mice were crossed with wild-type C57BL/6 mice to generate F1 offspring. heterzygotes F1 were crossed to get F2 mice. Peyer's patches of Trif- and Il1r1-deficient mice and their littermate wild-type controls (4 mice per group) were digested for 15 min at 37 °C in 0.5 mg/ml collagenase D and 10 μg/ml DNase I to generate a single-cell suspension. Live cells (Zombie NIR-negative) were sorted using a Cytek Aurora CS. One million cells from each sample were fixed with the ScaleBio Sample Fixation Kit. Fixed samples were stored at -80 °C until all samples were collected. Frozen fixed cells were thawed on ice and counted twice by hemocytometer. Using the ScaleBio Single Cell RNA Sequencing Kit v1.1 (Scale Biosciences), all 16 samples were prepared in one batch. Cells were loaded at 10,000 per well (6 wells per sample) to add RT barcodes as sample identifiers. Pooled cells were distributed across the 384-well plate for ligation barcode addition, then pooled again with 1,600 nuclei per well for second-strand synthesis. After tagmentation and indexing PCR, libraries were pooled (5 ul each), cleaned with 0.8X SPRIselect beads (Beckman Coulter B23317), and fragment size/concentration were quantified using High Sensitivity D5000 ScreenTape (Agilent).
GEO
GSM9693387_r1