PRJNA401805: Single-cell RNA-seq analysis of human CD14+ monocytes
概要: We performed single-cell RNA-seq on CD14+ monocytes isolated from the blood of healthy donors. Using the 10x chromium technology, we analyzed 425 and 431 cells from 2 individual donors.
项目整体设计: Peripheral Blood Mononuclear Cells (PBMC) were prepared by centrifugation on a Ficoll gradient. Blood CD14+ monocytes were isolated from healthy donors’ PBMC by positive selection using magnetic beads. Monocytes were 93-95% CD14+CD16- as assessed by flow cytometry. Cellular suspensions (1700 cells) were loaded on a 10X Chromium instrument (10X Genomics) according to manufacturer’s protocol.
| 生长方案: |
- |
| 处理方案: |
- |
| 提取方案: |
Single-cell RNA-Seq libraries were prepared using Chromium Single Cell 3’ v2 Reagent Kit (10X Genomics) according to manufacturer’s protocol |
| 建库方案: |
Indexed sequencing libraries were constructed using the reagents from the Chromium Single Cell 3’ v2 Reagent Kit, following these steps: (1) fragmentation, end repair and A-tailing; (2) size selection with SPRI select; (3) adaptor ligation; (4) post ligation cleanup with SPRI select; (5) sample index PCR and cleanup with SPRI select beads. |
| 分子类型: |
poly(A)+ RNA |
| 库的片段类型: |
PAIRED |
| 库的链类型: |
Forward |
| 测序平台: |
ILLUMINA |
| 测序仪型号: |
Illumina HiSeq 2500 |
| 链特异性: |
Specific |
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Condition Detail |
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加标(Spike-In) |
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Aryl Hydrocarbon Receptor Controls Monocyte Differentiation into Dendritic Cells versus Macrophages.
Immunity . 2017-09-01 [PMID:
28930664]