|
| Status |
Public on Oct 01, 2012 |
| Title |
HaCaT cells treated with S.aureus biofilm-conditioned media from cultures spiked with DMSO control, replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
Primary human keratinocytes were cultured from newborn foreskin.
|
| Organism |
Homo sapiens |
| Characteristics |
treatment: Conditioned medium from biofilm S. aureus cultures.
cell type: Human Foreskin Keratinocytes were isolated at the University of Washington.
|
| Treatment protocol |
After growing HaCaT cells to confluence, cells were treated for 4 hours at 37C, 5% CO2.
|
| Growth protocol |
HaCaT keratinocytes were cultured in 6 well plates using EpiLife growth medium supplemented with human keratinocyte growth supplement. Cells were maintained at 37C in a humidified, 5% CO2 atmosphere.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
After treatment, the medium was removed and RNA was isolated using an RNeasy minikit following the manufacturer’s instructions for adherent cells. Extracted RNA was then ethanol precipitated.
|
| Label |
biotin
|
| Label protocol |
Total RNA (500ng) was reverse transcribed, and amplified and biotin-labeled via in vitro transcription using the MessageAmp Premier kit
|
| |
|
| Hybridization protocol |
Labeled and fragmented cRNA was hybridized for 16 hr at 45C to the Affymetrix Human Genome U133A 2.0 GeneChip. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using an Affymetrix 7G scanner and GCOS version 1.4.
|
| Description |
BCM_b
|
| Data processing |
Microarray data were analyzed using FlexArray version 1.6. The Affymetrix CEL files were imported and normalized using GC-RMA.
Matrix files include scaled MAS5 (linked to the Series record) and GC-RMA (contained in the data table below) normalized (log2) values.
|
| |
|
| Submission date |
Oct 12, 2011 |
| Last update date |
Oct 01, 2012 |
| Contact name |
Kate McInnerney |
| E-mail(s) |
kmcinnerney@montana.edu
|
| Phone |
406-994-5666
|
| Fax |
406-994-4926
|
| URL |
http://cores.montana.edu
|
| Organization name |
Montana State University
|
| Department |
Microbiology
|
| Lab |
Functional Genomics Facility
|
| Street address |
109 Lewis Hall
|
| City |
Bozeman |
| State/province |
MT |
| ZIP/Postal code |
59717 |
| Country |
USA |
| |
|
| Platform ID |
GPL571 |
| Series (1) |
| GSE32920 |
Phevalin (aureusimine B) production by Staphylococcus aureus biofilm and impacts on human keratinocyte gene expression |
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