General Information
- Experiment ID
- OEX027261
- Experiment Name
- Old_c-Kit+_scRNA-seq
- Experiment Type
- Transcriptomic single cell
- Description
- c-Kit+ progenitor cells from old WT and KO-MDS mice were used for single cell RNA sequencing.
- Protocol
- BM cells separated from old WT and Setd2-MDS mice were stained with biotin-c-Kit antibody on ice for 20 minutes, then washed with MojoSort™ Buffer (480017, BioLegend) at 3,000rpm for 5 minutes. Subsequently, samples labelled with biotin-c-Kit antibody were isolated through MACS assay, then the c-Kit+ BM cells were purified with BD FACSAria™ III. After purification, the BD Rhapsody system (BD Biosciences) was used to capture the transcriptomic information of single cells. Then, a limiting dilution approach was performed to randomly sediment and capture the single cell. Beads with oligonucleotide barcodes were then added to reach a saturated state, ensuring that each cell can be paired with one bead. Cells subjected with beads were captured and used for scRNA-seq. Whole transcriptome libraries were constructed following the BD Rhapsody single-cell whole-transcriptome amplification workflow. Single-cell libraries were quantified using a High Sensitivity DNA chip (Agilent) on a Bioanalyzer 2200 and the Qubit High Sensitivity DNA Assay Kit (Thermo Fisher Scientific). The library was sequenced by HiSeq Xten (Illumina) on a 150 bp paired-end run.
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 PCR library_strategy RNA-Seq platform Illumina NovaSeq 6000 - Project
-
1
Project ID Project name Create Date OEP004811 Loss of Setd2 induces systemic inflammation via embryo-derived Kupffer-like cell differentiation to drive malignant transformation 2023-12-07 - Samples
-
1
Sample ID Sample Name Organism Tissue Create Date OES354767 Old_c-Kit+_scRNA-seq Mus musculus bone marrow 2024-01-26 - Runs
-
1
Run ID Run Name Sample Data Num Create Date OER458863 old_c-Kit+_scRNA-seq Old_c-Kit+_scRNA-seq 4 2024-01-26
Author Information
- Create Date
- 2024-01-26
- Last Modified
- 2024-02-27
- Submission
- Yuanliang Zhang
- Shanghai Institute of Hematology