|
| Status |
Public on Jul 07, 2010 |
| Title |
12R/11S - repeat 5 - rev. label - mAdbID:64104 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Sensitive Isolate - Cy5
|
| Organism |
Nakaseomyces glabratus |
| Characteristics |
developmental stage: log-phase culture
genotype/variation: WT
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol Extraction Protocol
Other: Total RNA was isolated from log phase culture of C. glabrata grown in YPD using Trizole (Invitrogen) and RNeasy MiniElute cleanup kit (Qiagen).
|
| Label |
cy5
|
| Label protocol |
Cy5 Labeling Protocol
Other: Thirty micrograms of total RNA were used for each labeling. The RNA was reversed transcribed to cDNA to incorporate the fluorescent Cy5-dUTP (GE Health Care).
|
| |
|
| Channel 2 |
| Source name |
Resistant Isolate - Cy3
|
| Organism |
Nakaseomyces glabratus |
| Characteristics |
developmental stage: log-phase culture
genotype/variation: CgPDR1 gain-of-function mutation
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol Extraction Protocol
Other: Total RNA was isolated from log phase culture of C. glabrata grown in YPD using Trizole (Invitrogen) and RNeasy MiniElute cleanup kit (Qiagen).
|
| Label |
cy3
|
| Label protocol |
Cy3 Labeling Protocol
Other: Thirty micrograms of total RNA were used for each labeling. The RNA was reversed transcribed to cDNA to incorporate the fluorescent Cy3-dUTP (GE Health Care).
|
| |
|
| |
| Hybridization protocol |
Hybridization Protocol
Other: Microarray hybridization was done at the NIH/NIAID/RTB core facility. The 70-mer oligo arrays were prehybridized at 42°C in prehybridization buffer (5X SSC, 1% BSA, 0.1% SDS) for 30-60 min and then hybridized to the labelled cDNA in 50 ul of hybridization buffer (25% formamide, 5X SSC, 0.2% SDS, 20 ug/ml Poly (dA40-60), 200 ug/ml Cot-1 DNA, 80 ug/ml yeast t-RNA) overnight at 42°C. The microarrays were washed three times in wash buffer A (1X SSC, 0.05% SDS) and washing buffer B (0.1X SSC).
|
| Scan protocol |
Creator: GenePix Pro 6.0.0.45
Scanner: GenePix 4000B [82425]
ScanPower: 100;; 100
LaserPower: 3.02;; 3.34
Temperature: 31
|
| Description |
mAdb experiment ID: 64104
|
| Data processing |
mAdb Data Processing
Calculation Method: Background correction was done using median intensity minus median background. Calculations were based on signals greater than 100 in both channels and excluding features designated as controls or flagged as bad or not found. Ratios were normalized using the 50th percentile (median).
|
| |
|
| Submission date |
Apr 15, 2010 |
| Last update date |
Apr 30, 2013 |
| Contact name |
Huei-Fung Tsai |
| E-mail(s) |
[email protected]
|
| Phone |
301-496-8993
|
| Fax |
301-480-0512
|
| Organization name |
NIH/NIAID
|
| Department |
LCID
|
| Lab |
CMS
|
| Street address |
10 Center Dr. Bldg. 10 Rm. 11N228
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL8174 |
| Series (1) |
| GSE21352 |
Azole Resistance Mechanism in Candida glabrata Oropharyngeal Isolates |
|
