|
Status |
Public on Jul 08, 2010 |
Title |
SC (time 1) Replicate 1 Dye Swap |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
Reference Pool SC
|
Organism |
Saccharomyces cerevisiae |
Characteristics |
reference: Reference Pool SC
|
Treatment protocol |
Add fluconazole (4ug/mL for Sc and Cg, 0.06125ug/mL for Kl) at time 0
|
Growth protocol |
Grow samples at OD600 0.05 to 0.2. Treat half of sample with drug. Take samples at 0, 1/3, 2/3, 1, 2 and 4 doubling times (both treated and untreated).
|
Extracted molecule |
total RNA |
Extraction protocol |
total RNA was extracted using hot phenol chloroform method. mRNA was isolated using Poly(A) purist kit (AM1916), Pool in equal amounts untreated samples for each replicate
|
Label |
Cy5
|
Label protocol |
Labeling performed using GE Healthcare Cyscribe First strand cDNA synthesis kit
|
|
|
Channel 2 |
Source name |
BY4741 time 1
|
Organism |
Saccharomyces cerevisiae |
Characteristics |
test: BY4741 doubling time: 1
|
Treatment protocol |
Add fluconazole (4ug/mL for Sc and Cg, 0.06125ug/mL for Kl) at time 0
|
Growth protocol |
Grow samples at OD600 0.05 to 0.2. Treat half of sample with drug. Take samples at 0, 1/3, 2/3, 1, 2 and 4 doubling times (both treated and untreated).
|
Extracted molecule |
total RNA |
Extraction protocol |
total RNA was extracted using hot phenol chloroform method. mRNA was isolated using Poly(A) purist kit (AM1916), Pool in equal amounts untreated samples for each replicate
|
Label |
Cy3
|
Label protocol |
Labeling performed using GE Healthcare Cyscribe First strand cDNA synthesis kit
|
|
|
|
Hybridization protocol |
Oligoarray control targets and hybridization buffer were added, and samples were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed sequentially
|
Scan protocol |
Arrays scanned using Genepix 4000A scanner and GenePix 6.0 software
|
Description |
dye swap replicate
|
Data processing |
Background subtraction and LOESS, missing values replaced by KNNimpute algorithm. Data was subsequently quantile normalized
|
|
|
Submission date |
Apr 16, 2009 |
Last update date |
Jul 08, 2010 |
Contact name |
Dwight Kuo |
Organization name |
UCSD
|
Department |
Bioengineering
|
Lab |
Trey Ideker
|
Street address |
9500 Gilman Dr.
|
City |
La Jolla |
State/province |
CA |
ZIP/Postal code |
92093 |
Country |
USA |
|
|
Platform ID |
GPL7293 |
Series (1) |
GSE15710 |
Evolutionary divergence in the fungal response to fluconazole revealed by soft clustering (S. cerevisiae, C. glabrata, K. lactis) |
|