|
| Status |
Public on Feb 02, 2018 |
| Title |
SRpP55 KD (siSR#2) exposed 2 |
| Sample type |
SRA |
| |
|
| Source name |
EndoC-beta H1 human cell line
|
| Organism |
Homo sapiens |
| Characteristics |
treatment: SRpP55 KD (siSR#2)
|
| Growth protocol |
Human insulin-producing EndoC-βH1 cells (Scharfmann et al., 2014) were grown on matrigel/fibronectin (100 and 2 μg/mL, respectively) coated plates and cultured in DMEM medium as previously described
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Total RNA was isolated from Maria using the RNeasy Mini kit (Qiagen, Venlo, The Netherlands) which favours purification of RNA molecules longer than 200 nucleotides. mRNA was purified from 1 μg total RNA using oligo (dT) beads, before it was fragmented and randomly primed for reverse transcription followed by second-strand synthesis to create ds cDNA fragments.
mRNA was purified from 1 µg total RNA using oligo (dT) beads, before it was fragmented and randomly primed for reverse transcription followed by second-strand synthesis to create ds cDNA fragments. The generated cDNA had undergone paired-end repair to convert overhangs into blunt ends. After 39- monoadenylation and adaptor ligation, cDNAs were purified on a 2% agarose gel and 200 basepair (bp) products were excised from the gel. Following gel digestion, purified cDNA was amplified by PCR using primers specific for the ligated adaptors. The generated libraries were submitted to quality control with the Agilent bio- analyzer 2100 (Agilent Technologies, Wokingham, UK) before sequencing. The RNA integrity number (RIN) values for all samples were 7.5 and above. 1 µl cDNA was loaded on an Agilent DNA chip (DNA-1000) to verify cDNA quality and quantity. Only libraries reaching satisfactory conditions were used for sequencing, on an Illumina HiSeq 2000 system
|
| |
|
| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina HiSeq 2000 |
| |
|
| Data processing |
Sequencing reads were mapped to the Homo sapiens genome (version GRCh37/hg19) using the program Tophat (http://ccb.jhu.edu/software/tophat/index.shtml) using default options the GENCODE 18 annotation of genes transcripts. The Tophat mapper reports all mappings and split-mappings.
GTF files: Mapped reads were used to quantify human transcripts from the GENCODE 18 annotation dataset, using the Flux Capacitor approach that deconvolves reads mapping to exonic regions shared by multiple transcripts by optimizing a system of linear equations and thus obtains a number of reads specifically assigned to each alternative spliceform (http://flux.sammeth.net). All genes and transcripts have been assigned a relative coverage rate as measured in RPKM units ("reads per kilobase per million mapped reads").
Genome_build: GRCh37/hg19
Supplementary_files_format_and_content: The processed data files are in standard transcript GTF format (for the full format description see http://www.ensembl.org/info/website/upload/gff.html). The attributes column contains informations on the transcript length, transcript abundance quantification RPKM (reads per kilobase per million mapped reads) and the number of read that mapped to it.
|
| |
|
| Submission date |
May 02, 2017 |
| Last update date |
May 15, 2019 |
| Contact name |
Jonas Juan Mateu |
| E-mail(s) |
mjuanmat@ulb.ac.be
|
| Phone |
003225556107
|
| Organization name |
Universite Libre de Bruxelles
|
| Department |
Faculty of Medecine
|
| Lab |
ULB Center for Diabetes Research
|
| Street address |
Route de Lennik 808 CP 618
|
| City |
Brussels |
| ZIP/Postal code |
1070 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL11154 |
| Series (1) |
| GSE98485 |
A SRp55-regulated alternative splicing network controls pancreatic beta cell survival and function |
|
| Relations |
| BioSample |
SAMN06883733 |
| SRA |
SRX2779456 |
