|
| Status |
Public on Dec 16, 2013 |
| Title |
mRNA-seq, RAD21cv HEK293, TEV treated, replicate two |
| Sample type |
SRA |
| |
|
| Source name |
mRNA-seq, RAD21cv HEK293, TEV treated
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HEK293T RAD21cv
|
| Treatment protocol |
A stably integrated transgene in the RAD21cv HEK293T cells was activated for 3 days with 2µg/mL of doxycycline. Cells were then transfected with a plasmid expressing TEV protease using Lipofectamine 2000 (Invitrogen) for 24 hours before harvesting
|
| Growth protocol |
Cells were grown in DMEM supplemented with 0.2mM L-glutamine, 100 U/mL penicillin, 100 mg/mL streptomycin, and 10% fetal calf serum at 37°C and 5% CO2
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Total RNA was extract using Trizol (Invitrogen) according to manufacturer's instructions. polyA RNA was isolated using Dynalbeads mRNA purification kit (Invitrogen) according to the manufacturer's instructions.
Strand Specific libraries were constructed according to Parkhomchuk et al. 2009, Nucleic Acids Res
|
| |
|
| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina HiSeq 2000 |
| |
|
| Data processing |
fastq: Illumina's HiSeq Control Software
Hi-C reads were aligned as single end reads using BWA with default parameters. Uniquely mapping reads were manually paired using in an in house pipeline. PCR duplicate reads were removed using Picard MarkDuplicates.
RNA-seq data were mapped as paired-end reads using tophat with the following parameters: -g 1 -p 12 --solexa1.3-quals --library-type fr-firststrand --segment-length 25 --bowtie1
RNA-seq bigWig files were generated through an in house pipeline that includes performing trimmed mean of M (TMM) normalization (Robinson et al. Genome Biology 2010)
ChIP-seq data were mapping with bowtie using the following parameters: -v 3 -m 1 --best --strata -S --time -p 8. Data was filtered for uniquely mapping reads. PCR duplicates were removed using Picard MarkDuplicates
ChIP-seq bigWig files were generated through an in house pipeline
Genome_build: hg18
Supplementary_files_format_and_content: ChIP-seq and RNA-seq processed files are in bigWig format. As the RNA-seq experiment was strand specific, bigWig files for both the positive and negative strands are included
Supplementary_files_format_and_content: Hi-C processed files are in a modified bed format. Each row lists the chromosome and the start and end coordiates of two interacting bins as well as the normalized interaction frequency between these two bins
Supplementary_files_format_and_content: Processed mRNA-seq data include information for each gene and its count and rpkm values. Each line includes chromosome, start position, end position, strand, gene name, read counts, and rpkm values
Genome Build:
TEV2.pos.norm.bw: hg18
TEV2.neg.norm.bw: hg18
TEV_r2.rpkm: hg18
|
| |
|
| Submission date |
Feb 12, 2013 |
| Last update date |
May 15, 2019 |
| Contact name |
Jesse R Dixon |
| E-mail(s) |
jedixon@salk.edu
|
| Organization name |
Salk Institute for Biological Studies
|
| Lab |
PBL-D
|
| Street address |
10010 N. Torrey Pines Rd.
|
| City |
La Jolla |
| State/province |
CA |
| ZIP/Postal code |
92037 |
| Country |
USA |
| |
|
| Platform ID |
GPL11154 |
| Series (1) |
| GSE44267 |
Cohesin and CTCF Differentially Affect the Chromatin Architecture and Gene Expression in Human Cells |
|
| Relations |
| SRA |
SRX235601 |
| BioSample |
SAMN01919646 |
