|
| Status |
Public on Sep 30, 2013 |
| Title |
NCTC8325 1980::ermB |
| Sample type |
RNA |
| |
|
| Source name |
S. aureus growth culture, OD600=2.0
|
| Organism |
Staphylococcus aureus subsp. aureus NCTC 8325 |
| Characteristics |
strain: SAOUHSC_01980-1981 mutated strain
|
| Treatment protocol |
In order to lysis the cell wall, cells were collected and incubated in 37°C with 10 mg/ml lysozyme and 40 ug/ml lysostaphin for 5 minutes. Then, cells were centrifuged and resuspended in TRIzol immediately.
|
| Growth protocol |
Overnight culture were diluted 1:100 in TSB and grow until OD600=2.0.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared using mirVana RNA Isolation Kit (Applied Biosystem p/n AM1556) following the manufacturer’s instruction.
|
| Label |
cy5
|
| Label protocol |
5 ug RNA was used to synthesize cDNA, and labeled with cy5. This process was accomplished using FairPlay III Microarray Labeling Kit (Aglilent p/n252012). Labeled cDNA was purified and quantified with NanoDrop Spectrophotometer (Thermo G2939A).
|
| |
|
| Hybridization protocol |
total of 600 ng cy5-labeled cDNA was used for hybridization sample using Gene Expression Hybridization Kit (Agilent p/n5188-4242) following the manufacturer’s instruction. 40 ul of the hybridization samples were added onto the microarray slide and hybridized in an Agilent hybridization oven to rotate at 10 rpm at 65 °C for 17 hours. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 8x15k array slides (Scan resolution 5 um, Dye channel is set to Red and Red PMT is set to 100%).
|
| Description |
Gene expression in SAOUHSC_01980-1981 mutated strain
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software (Agilent). Quantity normalization were analyzed with GENESPRING 11.5 (Agilent).
|
| |
|
| Submission date |
Sep 26, 2013 |
| Last update date |
Sep 30, 2013 |
| Contact name |
Haipeng Sun |
| Organization name |
University of Science and Technology of China
|
| Department |
School of Life Sciences
|
| Street address |
Huangshan Road
|
| City |
He Fei |
| State/province |
An Hui |
| ZIP/Postal code |
230027 |
| Country |
China |
| |
|
| Platform ID |
GPL17776 |
| Series (1) |
| GSE51197 |
Effect of different Two-component systems system deletion in S. aureus NCTC8325 |
|
