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Sample GSM2705441 Query DataSets for GSM2705441
Status Public on Jul 18, 2017
Title V76
Sample type SRA
 
Source name dorsal lateral prefrontal cortex (Brodmann Area 9)
Organism Homo sapiens
Characteristics diagnosis: DSM-IV major depressive disorder non-suicides (MDD)
age (yrs): 68
Sex: Female
rin: 5.3
pmi: 13
brain ph: 6
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 M17
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 SAMN07357750
SRA SRX3009325

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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