|
| Status |
Public on Feb 11, 2023 |
| Title |
SMARCA4_786O PBRM1WT_9hr_BTZ_ChIPSeq_rep2 |
| Sample type |
SRA |
| |
|
| Source name |
Cancer cell line
|
| Organism |
Homo sapiens |
| Characteristics |
source: 786-O (ATCC CRL1932)
protein: SMARCA4
treatment: bortezomib 5nM, 9hr
chip antibody: SMARCA4(AbCAM, ab110641, lot GR3375498-10)
|
| Treatment protocol |
Cells were treated with DMSO control or 5nM bortezomib for 9 hours prior to ChIP-seq. Cells were treated with DMSO control or 10uM SMARCA2/4 ATPase inhibitor (BRM014) for 24 hours prior to ChIPseq.
|
| Growth protocol |
Cells were grown in RPMI-1640 medium (Sigma) supplemented with 10% fetal bovine serum (Gibco). Cells were maintained in a 5% CO2-humidified incubator at 37°C.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
For each protein of interest, approximately 2x107 cells were cross-linked with 1% formaldehyde for 10 min at room temperature, and stopped by adding glycine to a final concentration of 0.2M. Chromatin was extracted and sonicated to ~500bp (Vibra cell, SONICS). The total volume of immunoprecipitation was 2 ml and the amount of antibody used was 20 µg. The input DNA was precleared with protein G Dynabeads (LifeTechnologies) for 2 hours at 4°C and then incubated with antibodies overnight at 4°C. Protein G beads were added the following day and mixture was nutated for 3 hours at 4°C. The beads were washed 6 times with wash buffer at room temperature.
At least 10 ng of the amplified DNA was used with NEBNext ChIP-Seq library prep reagent set (NEB). Each library was sequenced to an average depth of 30-50 million reads on Novaseq
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| |
|
| Library strategy |
ChIP-Seq |
| Library source |
genomic |
| Library selection |
ChIP |
| Instrument model |
Illumina NovaSeq 6000 |
| |
|
| Data processing |
Sequencing reads were mapped against human genome reference hg19 using the Burrows-Wheeler Aligner (BWA) (version 0.6.2) 'mem' algorithm. Only reads with mapQ >10 and with duplicate removed by rmdup were used in the subsequent analysis
Significant peaks were called using MACS2 24 (q-value < 0.01).
Assembly: hg19
Supplementary files format and content: ChIP-seq peaks are in macs2 narrowPeak format, including 10 columns: "chromosome" "start coordinate" "end coordinate" "name" "score" "strand" "signalValue" "pValue" "qValue" "peak"
|
| |
|
| Submission date |
Jan 05, 2023 |
| Last update date |
Feb 11, 2023 |
| Contact name |
Xiaosai Yao |
| E-mail(s) |
xiaosai.yao@gmail.com, yao.xiaosai@gene.com
|
| Phone |
+65 68088271
|
| Organization name |
Genome Institute of Singapore
|
| Street address |
60 Biopolis Street, Genome, #02-01
|
| City |
Singapore |
| ZIP/Postal code |
138672 |
| Country |
Singapore |
| |
|
| Platform ID |
GPL24676 |
| Series (2) |
| GSE152735 |
Genome-wide chromatin profiles of PBRM1 |
| GSE222244 |
Genome-wide chromatin profiles of PBRM1 Drug treatment [ChIP-Seq] |
|
| Relations |
| BioSample |
SAMN32602237 |
| SRA |
SRX18945308 |
