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Sample GSM5526318 Query DataSets for GSM5526318
Status Public on Aug 20, 2021
Title patient 1 tongue rep 2
Sample type SRA
 
Source name tongue autopsy tissue
Organism Homo sapiens
Characteristics replicate type: technical replicate 2
viral rna detected?: no
Extracted molecule total RNA
Extraction protocol Total RNA was extracted from the autopsy tissues by first flash freezing each tissue in liquid nitrogen and then using the CryoPREP Tissue Disruption System (Covaris) to pulverize the tissue into a fine powder. The powder was then immediately resuspended in cell lysis buffer and RNA was extracted using the ReliaPrep RNA Tissue Miniprep System (Promega, #Z6112) by following the manufacturer’s instructions. A DNase digestion was also performed during the RNA extraction workflow to remove any potential contamination from carryover genomic DNA (reagents included in the kit). The extracted RNA was then purified using 1.8X RNAClean XP beads (Beckman Coulter) to remove any molecular impurities.
Stranded total RNA libraries were generated according to the manufacturer's instructions, using the TruSeq Stranded Total RNA Gold kit for Illumina sequencing
RNAseq, 2x100 PE reads, Illumina NovaSeq 6000
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina NovaSeq 6000
 
Description s6-tongue-rep2
Data processing The raw RNAseq data was aligned to the hg38 genome using the STAR(v2.7.3a) mapper.
Aligned data were annotated using GENCODE v35 gene annotations and counted using featureCounts(v1.6.3).
Genes with fewer than 50 counts across all samples were excluded from the analysis, and the counts were subsequently log2-transformed for differential gene expression analysis using DESeq2. A pairwise comparison between infected tissues (lung, nasopharynx) and uninfected tissues (olfactory bulb, prefrontal cortex, oropharynx, salivary gland, tongue, heart, liver, kidney) with an adjusted p-value < 0.05 for differentially expressed (DE) genes.
Gene Set Enrichment Analysis (clusterProfiler) was performed to probe for Gene Ontology (GO) categories that were significantly overrepresented and statistically different in infected versus non-infected tissues.
Genome_build: hg38
Supplementary_files_format_and_content: featureCount tsv file summarizes raw counts from featureCounts for each sample.
Supplementary_files_format_and_content: DESeq csv file contains DE genes found by DESeq2.
Supplementary_files_format_and_content: GSEA csv file contains GESA results for GO categories.
 
Submission date Aug 17, 2021
Last update date Aug 20, 2021
Contact name Robert Sebra
Organization name Icahn School of Medicine at Mount Sinai
Department Genetics and Genomic Sciences
Street address One Gustave L. Levy Place
City New York
State/province NY
ZIP/Postal code 10029
Country USA
 
Platform ID GPL24676
Series (2)
GSE182297 Molecular Profiling of COVID-19 Autopsies Uncovers Novel Disease Mechanisms [RNA-Seq]
GSE182299 Molecular Profiling of COVID-19 Autopsies Uncovers Novel Disease Mechanisms
Relations
BioSample SAMN20839516
SRA SRX11802668

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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