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Single cells from the small intestine were obtained by digestion with papain (0.8 U/ml, Sigma, P3125), DNase i (50 U/ml, Coolaber, CD4871), FBS (20 μl/ml, Gibco), collagenase i (31.25 U/ml, Sigma, C0130), and collagenase iv (31.25 U/ml, Sigma, C5138). Single-cell suspensions of freshly isolated cells were then resuspended in PBS containing 0.04% ultrapure BSA. scRNA-seq libraries were prepared using Chromium Single Cell 3' Reagent Kits v2 (10x Genomics) according to the manufacturer’s instructions. The target cell recovery for each library was 7,000. Generated libraries were sequenced on an Illumina HiSeq2500 system (Beijing Institutes of Life Science, Chinese Academy of Science). |
RNA-Seq |
TRANSCRIPTOMIC SINGLE CELL |
RANDOM |
PAIRED
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