|
| Status |
Public on Apr 05, 2012 |
| Title |
C3 veg vs C5 sex 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Mycelia
|
| Organism |
Neurospora intermedia |
| Characteristics |
type: vegetative mycelium
|
| Treatment protocol |
Constant Darkness
|
| Growth protocol |
Vegetative Nutrient Medium (Vogel's Medium)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
2 µg of mRNA were primed with .5 µg of Oligo-d(T) primer at 70°C for 10min, then reversed transcribed at 42°C for 2h in the presence of 40 U SuperScript II RTase (Invitrogen), and .8ul Amino-allyl dUTP/dNTP made from Sigma in mg aliquotes and dissolved in 20µl H2O-(DEPC) and mixed with 30µl of 100mM dATP, dTTP, dGTP, dCTP and 12µl 100mM dTTP; 25 µM Cy3/Cy5-label
|
| |
|
| Channel 2 |
| Source name |
Mycelia
|
| Organism |
Neurospora intermedia |
| Characteristics |
type: sexual mycelium
|
| Treatment protocol |
Constant Darkness
|
| Growth protocol |
Vegetative Nutrient Medium (Vogel's Medium)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
2 µg of mRNA were primed with .5 µg of Oligo-d(T) primer at 70°C for 10min, then reversed transcribed at 42°C for 2h in the presence of 40 U SuperScript II RTase (Invitrogen), and .8ul Amino-allyl dUTP/dNTP made from Sigma in mg aliquotes and dissolved in 20µl H2O-(DEPC) and mixed with 30µl of 100mM dATP, dTTP, dGTP, dCTP and 12µl 100mM dTTP; 25 µM Cy3/Cy5-label
|
| |
|
| |
| Hybridization protocol |
Samples were applied to microarrays enclosed in hybridization chambers (53°C water bath). After hybridization, slides were washed sequentially
|
| Scan protocol |
Hybridized microarrays were scanned and gridded with a GenePix 4000B microarray scanner (Axon Instruments, Foster City, CA)
|
| Description |
veg_mc vs sex_mc
dye-swap
|
| Data processing |
Normalized by background-subtracted mean-by-mean normalization of well measured spots, and statistically analyzed using a Bayesian analysis of gene expression levels with BAGEL software.
|
| |
|
| Submission date |
Apr 04, 2012 |
| Last update date |
Apr 05, 2012 |
| Contact name |
Nicklas Samils |
| E-mail(s) |
[email protected]
|
| Organization name |
Swedish University of Agricultural Sciences
|
| Department |
Forest Mycology and Plant Pathology
|
| Street address |
Almas allé 5
|
| City |
Uppsala |
| ZIP/Postal code |
SE75007 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL10640 |
| Series (1) |
| GSE37034 |
Rapid evolution in fungal reproductive genes as revealed by evolutionary analyses of expressed sequence tags in Neurospora |
|
