|
| Status |
Public on Jan 26, 2011 |
| Title |
a4200 vs A2489 [Veg] |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Mycelia
|
| Organism |
Neurospora crassa |
| Characteristics |
mating type: Mat-a FGSC#4200
growth medium: Vegetative Nutrient Medium (Vogel's Medium)
|
| 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 crassa |
| Characteristics |
mating type: Mat-A FGSC#2489
growth medium: Vegetative Nutrient Medium (Vogel's Medium)
|
| 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 |
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 |
Jan 21, 2011 |
| Last update date |
Jan 26, 2011 |
| Contact name |
Nicklas Samils |
| E-mail(s) |
nicklas.samils@ebc.uu.se
|
| Organization name |
Uppsala University
|
| Department |
Evolutionary Biology
|
| Street address |
Norbyägen 18D
|
| City |
Uppsala |
| ZIP/Postal code |
75236 Uppsala |
| Country |
Sweden |
| |
|
| Platform ID |
GPL10640 |
| Series (1) |
| GSE26798 |
Mating-type specific gene expression in the filamentous ascomycete Neurospora tetrasperma |
|
