|
| Status |
Public on Sep 08, 2015 |
| Title |
host-Normal-PMNs-time-24-rep2 |
| Sample type |
RNA |
| |
|
| Source name |
host-Normal-PMNs-time-24-rep2
|
| Organism |
Homo sapiens |
| Characteristics |
strain: normal-PMNs--not infected with Gb
infection: not infected with Gb
cell type: Polymorphonuclear leukocytes
disease state: normal
time: 24 h
|
| Treatment protocol |
Cultures contained 1 ml of 10% subject autologous serum in RPMI 1640 (Gibco) supplemented with 25 mM HEPES pH 7.2 with or without 1 × 107 PMN with or without 1 × 108 G. bethesdensis.
|
| Growth protocol |
PATHOGEN-G. bethesdensis was grown to mid-log phase at 37C in YPG medium : HOST-Serum and Percoll-isolated PMN were isolated from venous blood
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RLT cell lysates were homogenized using QIAshredder system and neutrophil and bacterial RNA were simultaneously extracted using an AllPrep DNA/RNA 96 Kit (Qiagen) with on-column DNase I treatment
|
| Label |
biotin
|
| Label protocol |
PATHOGEN-Life Technologies (Carlsbad, CA) MessageAmp™ II-Bacteria kit was used as recommended to amplify an average of 0.3 mg of G. bethesdensis and human neutrophil RNAs; HOST Five ng of RNA sample was concentrated to 5 μL and processed according to manufacturer’s instructions using WT-Ovation™ Pico system RNA Amplification System (Nugen Inc., San Carlos, CA).
|
| |
|
| Hybridization protocol |
PATHOGEN-Samples were hybridized onto custom Affymetrix GeneChip; HOST-samples were hybridized onto Affymetrix human U133 plus 2.0 arrays according to manufacturer's recommendations
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 7GPlus.
|
| Description |
Gene expression data from_host-Normal-PMNs-time-24-rep2
|
| Data processing |
The data were analyzed with EC 1.1 and scaled to 500
|
| |
|
| Submission date |
Mar 12, 2014 |
| Last update date |
Sep 08, 2015 |
| Contact name |
Dan Sturdevant |
| E-mail(s) |
dsturdevant@niaid.nih.gov
|
| Phone |
4063639248
|
| Organization name |
NIH
|
| Department |
NIAID
|
| Lab |
RTS
|
| Street address |
903 S 4th street
|
| City |
Hamilton |
| State/province |
MT |
| ZIP/Postal code |
59840 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE55849 |
Simultaneous Host-Pathogen transcriptome analysis during Granulibacter bethesdensis infection of normal and chronic granulomatous disease neutrophils |
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