PRJNA464435: Single nucleus inDrops to profile adult human kidney
概要: Single nucleus inDrops to profile adult human kidney
项目整体设计: Single nuclei were isolated from adult human kidney and subjected to inDrops single cell RNA-seq
| 生长方案: |
- |
| 处理方案: |
- |
| 提取方案: |
Nuclei were isolated with Nuclei EZ Lysis buffer (Sigma #NUC-101) supplemented with protease inhibitor (Roche #5892791001) and RNase inhibitor (Promega #N2615, Life Technologies #AM2696). Samples were cut into <2 mm pieces and homogenized using a Dounce homogenizer (Kimble Chase #885302-0002) in 2ml of ice-cold Nuclei EZ Lysis buffer and incubated on ice for 5 min with an additional 2ml of lysis buffer. The homogenate was filtered through a 40-碌m cell strainer (pluriSelect #43-50040-51) and then centrifuged at 500 x for 5 min at 4 潞C. The pellet was resuspended and washed with 4 ml of the buffer and incubated on ice for 5 min. After another centrifugation, the pellet was resuspended with Nuclei Suspension Buffer (1x PBS, 0.07% BSA, 0.1% RNase inhibitor), filtered through a 20-碌m cell strainer (pluriSelect 43-50020-50) and counted. We used the inDrops protocol developed by Klein et al to generate single nucleus transcriptome.RNA libraries were prepared for sequencing using standard inDrops procedure |
| 建库方案: |
- |
| 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 |
| 数据来源 |
GEN样本编号 |
GEN数据集编号 |
系列编号 |
项目编号 |
样本编号 |
样本名称 |
生物样本编号 |
样本访问号 |
实验访问号 |
释放时间 |
提交时间 |
最后更新时间 |
物种 |
种族 |
族裔 |
年龄 |
年龄单位 |
性别 |
来源名称 |
组织 |
细胞类型 |
细胞亚型 |
细胞系 |
疾病 |
疾病状态 |
发育阶段 |
突变/变异 |
表型 |
Condition Detail |
生长方案 |
处理方案 |
提取方案 |
建库方案 |
分子类型 |
库的片段类型 |
链特异性 |
库的链类型 |
加标(Spike-In) |
测序方法 |
测序平台 |
测序仪型号 |
细胞数 |
测序片段数 |
碱基数 |
平均测序片段长度_1 |
平均测序片段长度_2 |
唯一比对率 |
多重比对率 |
覆盖度 |
Single-Cell Transcriptomics of a Human Kidney Allograft Biopsy Specimen Defines a Diverse Inflammatory Response.
Journal of the American Society of Nephrology : JASN . 2018-07-06 [PMID:
29980650]