|
|
GEO help: Mouse over screen elements for information. |
|
Status |
Public on Jul 18, 2017 |
Title |
V71 |
Sample type |
SRA |
|
|
Source name |
dorsal lateral prefrontal cortex (Brodmann Area 9)
|
Organism |
Homo sapiens |
Characteristics |
diagnosis: non-psychiatric controls (CON) age (yrs): 51 Sex: Male rin: 6.8 pmi: 17 brain ph: 6.16 tissue: dorsal lateral prefrontal cortex (Brodmann Area 9)
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was extracted from dorsal prefrontal cortex (BA9) as previously described (Sibille E, Arango V, Galfalvy HC, Pavlidis P, Erraji-Benchekroun L, Ellis SP, et al. Gene expression profiling of depression and suicide in human prefrontal cortex. Neuropsychopharmacology. 2004;29(2):351–61) and according to the guidelines recommended by the NIH Roadmap Epigenomics Mapping Consortium (REMC). We used the Ambion’s mirVana miRNA isolation kit (#. AM1560, Life Technologies, Carlsbad, CA, USA) to isolate total RNA from postmortem brain tissue (Brodmann Area 9). The coding RNA library was generated using the TruSeq Stranded Total RNA Sample Prep kit (Illumina, San Diego, CA, USA) which includes rRNA depletion and chemical fragmentation. Paired-end, strand specific sequencing for total RNA was performed on Illumina HiSeq 2500 with 100 bp read lengths.
|
|
|
Library strategy |
miRNA-Seq |
Library source |
transcriptomic |
Library selection |
size fractionation |
Instrument model |
Illumina MiSeq |
|
|
Description |
M12
|
Data processing |
For each clinical sample, raw RNA-seq reads were aligned and mapped to the Ensembl GRCh37 human reference genome and assembled using Tophat v2.0.9 resulting in BAM files for each of 59 samples. Between 17,000,000 and 57,000,000 reads were obtained for each sample, and ~75–90% of reads were successfully mapped to the genome for each sample. Of these reads, ~10–15% aligned to multiple genomic loci. For each BAM file resulting from Tophat2, read counts per gene were summarized using the ‘summarizeOverlaps’ (union mode) function in the GenomicAlignments R library and a transcript database derived from the GRCh37 human genome assembly. Read counts per gene were normalized and written to csv file using the 'counts' function in DeSeq2 v1.6.3 For micro RNA samples, sequence adaptors were first trimmed i.e. using the command "cutadapt -b GATCGTCGGACTGTAGAACTCTGAAC -b TGGAATTCTCGGGTGCCAAGG", the resulting FastQ files were then converted to FASTA format and processed according to the Small RNA Analysis pipeline in CLC Genomics Workbench version 7.5 (http://www.clcbio.com/products/clc-genomics-workbench/ ) using default settings (i.e. mature length variants were allowed 2 additional or missing upstream and downstream base-pairs and 2 mismatched base-pairs, and alignment was strand-specific). Unique reads were extracted and counted and subsequently merged and annotated according to known miRNA species included in miRBase Release 21. Total counts for each miRNA (1,353 unique annotations with positive counts for one or more samples) were divided by the total library size to arrive at an expression value for each sample. Genome_build: GRCh37 human genome assembly Supplementary_files_format_and_content: counts_output.csv: Rows are gene ensemble IDs, columns are sample IDs
|
|
|
Submission date |
Jul 17, 2017 |
Last update date |
May 15, 2019 |
Contact name |
Spiro Pantazatos |
E-mail(s) |
spp2101@columbia.edu
|
Organization name |
Columbia University
|
Department |
Psychiatry
|
Street address |
1051 Riverside Dr.
|
City |
New York |
State/province |
New York |
ZIP/Postal code |
10032 |
Country |
USA |
|
|
Platform ID |
GPL15520 |
Series (1) |
GSE101521 |
Whole-transcriptome brain expression and exon-usage profiling in major depression and suicide |
|
Relations |
BioSample |
SAMN07357797 |
SRA |
SRX3009320 |
Supplementary data files not provided |
SRA Run Selector |
Raw data are available in SRA |
Processed data are available on Series record |
|
|
|
|
|