|
Status |
Public on Aug 25, 2009 |
Title |
∆clr4 cells |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
log. growing cells
|
Organism |
Schizosaccharomyces pombe |
Characteristics |
genotype: ∆clr4 cells
|
Growth protocol |
Standard conditions were used to produce logarithmically growing cultures in rich media (YEA) at 30˚C.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted using Master Pure Yeast RNA purification kit (Epicenter) according manufacturer's instructions.
|
Label |
Cy5
|
Label protocol |
Ribosomal RNA was depleted with RiboMinus kit (Invitrogen) and total RNA reverse-transcribed and labeled using SuperScript Indirect cDNA labeling kit (Invitrogen).
|
|
|
Channel 2 |
Source name |
log. growing cells
|
Organism |
Schizosaccharomyces pombe |
Characteristics |
genotype: wild-type cells
|
Growth protocol |
Standard conditions were used to produce logarithmically growing cultures in rich media (YEA) at 30˚C.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted using Master Pure Yeast RNA purification kit (Epicenter) according manufacturer's instructions.
|
Label |
Cy3
|
Label protocol |
Ribosomal RNA was depleted with RiboMinus kit (Invitrogen) and total RNA reverse-transcribed and labeled using SuperScript Indirect cDNA labeling kit (Invitrogen).
|
|
|
|
Hybridization protocol |
Equal amounts of Cy5-labeled mutant cDNA and Cy3-labeled wild-type cDNA were mixed and combined with human Cot1DNA, Agilent Blocking Agent and Agilent Hybridization buffer, and hybridized to high-density microarrays in Agilent SureHyb hybridization chamber for 24 hours at 65˚C, 10 rpm. After hybridization, slides were washed according to Agilent protocol.
|
Scan protocol |
Scanned on an Agilent G2505B scanner.
|
Description |
n/a
|
Data processing |
Data were extracted using Agilent Feature Extraction Software (GE2-v5_95_Feb07 protocol) and subjected to combined rank consistency filtering with LOWES intensity normalization. Background signal was estimated as a median processed signal of 152 oligonucleotides with no homology to S. pombe genome.
|
|
|
Submission date |
Jul 23, 2009 |
Last update date |
Aug 25, 2009 |
Contact name |
Shiv Grewal |
Phone |
2407607553
|
Organization name |
NCI
|
Department |
LBMB
|
Lab |
Shiv Grewal
|
Street address |
NCI bldg 37 Rm 6068 9000 Rockville Pike
|
City |
Bethesda |
State/province |
MD |
ZIP/Postal code |
20892 |
Country |
USA |
|
|
Platform ID |
GPL8908 |
Series (1) |
GSE17271 |
Histone H2A.Z cooperates with RNAi and heterochromatin factors to suppress antisense RNAs |
|