General Information
- Sample ID
- OES319152
- Sample Name
- HC1-H3K4me3_CHIP-seq
- Description
- CD4+T cells of healthy control
- Subject type
- Human
Sample information
- Organism
- Homo sapiens
- Tissue
- PBMC
- Protocol
- A sum of 20 mL of peripheral blood from each patient and healthy controls was preserved with heparin. Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation (GE Healthcare, Switzerland) , and then CD4+T cell are sorted by positive selection using CD4 beads (Miltenyi, Germany) respectively. The purity of CD4+ T cells was generally >95%.
- Projects
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1
Project ID Project name Submitter Date OEP004403 H3K4me3 ChIP-seq of CD4+T cell from SLE patients and HC Feng, Delong 2023-08-11 - Experiments
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1
Experiment ID Experiment name Experiment type Experiment protocol Date OEX024597 H3K4me3 ChIP-seq for of CD4+T cells form SLE patient and healthy control Genomic ChIP-seq was performed as previously described. Briefly, 2-5 million CD4+ T cells were lysed in cell lysis buffer for 5 min on ice. Cell were pelleted and resuspened with 200 μl of digestion buffer. Digestion was then performed by adding micrococcal nuclease and incubation for 5 min at 37 °C. The reaction was stopped by EDTA-EGTA buffer . After centrifuge for 15 min at 13000rpm, 1/10 of the supernatant were taken as Input sample, others supernatant was dilute into a CHIP-IP buffer. Chromatin was then incubated overnight at 4 °C with anti-trimethylated H3K4 antibody . Protein G magnetic beads were added for an additional 4 h of incubation. Immunoprecipitates were washed vigorously and DNA was purified with PCR purification kit. 1ng-50ng H3K4me3 ChIP'ed DNA or input DNA was used for ChIP-seq library construction via TruePrep DNA Library Prep Kit V2 for Illumina. After PCR step, the DNA libraries were purified and size selection by 0.5× and 0.25× DNA beads. ChIP-seq libraries were sequenced via paired-end 150 bp reads on an Illumina platform. 2023-08-12 - Runs
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1
Run ID Run Name Project Experiment Sample Data Date OER427260 HC1_H3K4me3_fq OEP004403 OEX024597 OES319152 2 2023-08-12
Author Information
- Create Date
- 2023-08-12
- Last Modified
- 2023-08-12
- Submission
- Delong Feng,The Second Xiangya hospital of Central South University