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| Status |
Public on Jun 27, 2019 |
| Title |
PBMCs, 0HC1 |
| Sample type |
SRA |
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| Source name |
canine peripheral blood mononuclear cells
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| Organism |
Canis lupus familiaris |
| Characteristics |
tissue: whole blood
cell type: PBMC
age: 10 months old
infection: control
time: 0h
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| Extracted molecule |
total RNA |
| Extraction protocol |
total RNA was extracted from the cells using the RNeasy Mini Kit (Qiagen, Germany) according to the manufacturer’s instruction.
the construction of library was performed using QuantSeq 3' mRNA-Seq Library Prep Kit (Lexogen, Inc., Austria) according to the manufacturer's instructions. In brief, each 500ng total RNA were prepared and an oligo dT primer containing an Illumina compatible sequence at its 5 end was hybridized to the RNA and reverse transcri ption was performed. After degradation of the RNA template, second strand synthesis was initiated by a random primer containing an Illumina compatible linker sequence at its 5 end. T he double stranded library was purified by using magnetic beads to remove all reaction components. The library was amplified to add the complete adapter sequences required for cluster generation. The finished library is purified from PCR components. High throughput sequencing was performed as single end 75 sequencing using Next Seq 500 (Illumina, Inc., USA).
QuantSeq
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| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina NextSeq 500 |
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| Data processing |
QuantSeq 3' mRNA Seq reads were aligned using Bowtie2 (Langmead and Salzberg, 2012).
Bowtie2 indices were either generated from genome assembly sequence or the representative transcript sequences for aligning to the genome and transcriptome.
The alignment file was used for assembling transcripts, estimating their abundances and detecting differential expression of genes.
Differentially expressed gene were determined based on counts from unique and multiple alignments using coverage in Bedtools (Quinlan AR, 2010).
The RC (Read Count) data were processed based on quantile normalization method using EdgeR within R (R development Core Team, 2016) using Bioconductor (Gentleman et al., 2004).
Genome_build: canFam3
Supplementary_files_format_and_content: abundance measurement
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| Submission date |
Jun 26, 2019 |
| Last update date |
Jun 30, 2019 |
| Contact name |
Suji Kim |
| E-mail(s) |
sujiksj43@gmail.com
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| Organization name |
Seoul National University
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| Department |
Veterinary Medicine
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| Lab |
Infectious Disease
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| Street address |
1, Gwanak-ro
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| City |
Seoul |
| State/province |
Gwanak-gu |
| ZIP/Postal code |
ASI/KR/KS013 |
| Country |
South Korea |
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| Platform ID |
GPL21400 |
| Series (1) |
| GSE133326 |
Global gene networks in canine peripheral blood mononuclear cells infected with Mycobacterium avium subsp.hominissuis |
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| Relations |
| BioSample |
SAMN12139329 |
| SRA |
SRX6367502 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSM3905777_0h-0C1.txt.gz |
150.8 Kb |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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