General Information
- Experiment ID
- OEX022989
- Experiment Name
- OEX_Shen_2304051419
- Experiment Type
- Transcriptomic
- Description
- Sample preparation and single cell RNA-seq library construction and sequencing
- Protocol
- The BALF was passed through a 100 μm filter (Falcon,3523260) and centrifugated (400g, 10 min, 4℃). Pelleted cells were resuspended in red blood cell lysis buffer (Solarbio, R1010) and incubated for 2 min, then passed through a 40 μm filter (Falcon,3523240), collected by centrifugation (400g, 10 min, 4℃) and resuspended in PBS (BI, 02-024-1ACS) containing 0.04% BSA (Sigma, B2064). Cells were manually counted by Trypan blue (Thermo, T10282) and AO-PI (LUNA, D23001) after each centrifugation (400g, 10 min, 4℃) and resuspended. Single cells were processed using Chromium Controller (10X Genomics) according to the manufacturer's protocol. By using Chromium Next GEM Single Cell 3' Kit v3.1 (10x Genomics, 1000268) and Chromium Next GEM Chip G Single Cell Kit (10x Genomics, 1000120), we performed single cell 3' gene expression profiling. The cell suspension was loaded onto the Chromium single cell controller (10x Genomics) to generate single-cell gel beads in the emulsion according to the manufacturer's protocol. Captured cells were lysed and the released RNA were barcoded through reverse transcription in individual GEMs. Cell-barcoded 3' gene expression libraries were sequenced on an lllumina NovaSeq6000 system by Shanghai Biochip Co., Ltd., Shanghai, China.
Experiment information
- Attributes
-
library_layout Paired mate_pair n/a insertion_size n/a number_of_reads_sequenced n/a library_name n/a library_selection PolyA library_strategy RNA-Seq platform Illumina NovaSeq 6000 - Project
-
1
Project ID Project name Create Date OEP003526 Single-cell transcriptomics of human bronchoalveolar lavage fluid from sepsis patients and healthy controls 2022-07-15 - Samples
-
1
Sample ID Sample Name Organism Tissue Create Date OES304322 Patient and Healthy samples Homo sapiens bronchoalveolar lavage 2023-04-05 - Runs
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1
Run ID Run Name Sample Data Num Create Date OER394040 OER_Shen_2304120941 Patient and Healthy samples 12 2023-04-12
Author Information
- Create Date
- 2023-04-05
- Last Modified
- 2023-07-20
- Submission
- Shen Rong
- Nanfang hospital