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Sample GSM95324 Query DataSets for GSM95324
Status Public on Feb 27, 2006
Title Brain_Wake_4hr_1
Sample type RNA
 
Source name Whole Brain_Wake_4hr
Organism Drosophila melanogaster
Characteristics Strain: Canton-S
Gender: Female
Age: 5 days
Tissue: Whole Brain
Time of Sacrifice: ZT14
Condition: Undisturbed Control
Treatment protocol Animals were sacrificed by immersion in >95°C distilled water, removed from water and stored at -70 ° C until tissue dissection performed within 24 hours of sacrifice. Brains were removed from the head and cleared of surrounding tissues using forceps under a dissecting microscope under ice cold insect ringers (130 mM NaCl, 4.7 mM KCl, 1.9 mM CaCl2) and immediately placed in Trizol® Reagent (U.S.Patent No.5,346,994).
Growth protocol Females were collected as newly eclosed virgins and kept in populations of 50 flies at 25°C under 12 hour light: 12 hour dark conditions in vials containing standard cornmeal media until time of sacrifice.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted from the pools of 139 (min) to 193 (max) brains using TRIZOL Reagent (U.S.Patent No.5,346,994) following the manufactrurers instructions (Invitrogen)
Label Biotin
Label protocol All protocols were conducted as described in the Affymetrix GeneChip Expression Analysis Technical Manual. Briefly, 5.0 ug of total RNA was converted to first-strand cDNA using Superscript II reverse transcriptase primed by a poly(T) oligomer that incorporated the T7 promoter. Second-strand cDNA synthesis was followed by in vitro transcription for linear amplification of each transcript and incorporation of biotinylated CTP and UTP. Performed by the staff at the Penn Microarray Facility http://www.med.upenn.edu/microarr/index.htm
 
Hybridization protocol The cRNA products were fragmented to 200 nucleotides or less, heated at 99ºC for 5 min and hybridized for 16 h at 45ºC to 1 microarray. The microarrays were then washed at low (6X SSPE) and high (100mM MES, 0.1M NaCl) stringency and stained with streptavidin-phycoerythrin. Fluorescence was amplified by adding biotinylated anti-streptavidin and an additional aliquot of streptavidin-phycoerythrin stain. Performed by the staff at the Penn Microarray Facility http://www.med.upenn.edu/microarr/index.htm
Scan protocol A confocal scanner was used to collect fluorescence signal at 3um resolution after excitation at 570 nm. The average signal from two sequential scans was calculated for each microarray feature. Performed by the staff at the Penn Microarray Facility http://www.med.upenn.edu/microarr/index.htm
Description This is one of 3 timepoints (4 Hour wake) in a study of the effects of stimulation during the active period on gene expression in the Drosophila brain.
Data processing Ten time groups were used in this study, each with three biological replicates. Seven time groups were used to determine differential gene expression between normal and sleep-deprived flies while three time groups were used as an experimental control to test for differential gene expression due to the effect of mechanical stimulation. Probe intensity data from all 30 arrays was read into the R software environment (http://www.R-project.org) directly from .CEL files using the R/AFFY package (Gautier et al., 2004). R/AFFY was also used to extract and manipulate probe level data to assess data quality and to create expression summary measures. Normalization was carried out using the robust multi-array average (RMA) method (Irizarry et al., 2003) on all probe intensity data sets together in order to form one expression measure for each gene on each array. Briefly, the RMA method adjusted the background of perfect match (PM) probes, applied a quantile normalization of the corrected PM values, and calculated final expression measures using the median polish algorithm.
 
Submission date Feb 02, 2006
Last update date Feb 23, 2006
Contact name John E Zimmerman
E-mail(s) johnez@mail.med.upenn.edu
Phone 215-746-4804
Fax 215-746-4813
URL http://www.uphs.upenn.edu/sleepctr/centerforsleep.htm
Organization name University of Pennsylvania School of Medicine
Department Center for Sleep & Respiratory Neurobiology
Lab Dr. Allan I. Pack
Street address 125 South 31st Street
City Philadelphia
State/province PA
ZIP/Postal code 19104
Country USA
 
Platform ID GPL72
Series (1)
GSE4174 Effect of Sleep Deprivation on the Whole Brain of Drosophila

Data table header descriptions
ID_REF
VALUE RMA of raw signal data from .CEL files
ABS_CALL Affymetrix Microarray Suite 5.0 detection call
DETECTION P-VALUE Affymetrix Microarray Suite 5.0 detection P-value

Data table
ID_REF VALUE ABS_CALL DETECTION P-VALUE
141200_at 3.384729 A 0.099061
141201_at 5.586031 A 0.562354
141202_at 6.668029 P 0.001755
141203_at 5.269906 P 0.000613
141204_at 4.458014 P 0.017732
141205_at 5.626348 P 0.000944
141206_at 6.829332 P 0.000491
141207_at 4.206061 P 0.000491
141208_at 5.324542 P 0.001755
141209_at 7.4516 P 0.002607
141210_at 3.425811 A 0.181343
141211_at 4.151536 P 0.003815
141212_at 8.023616 P 0.000613
141213_at 2.855502 A 0.181343
141214_at 7.942347 P 0.000491
141215_at 6.880328 P 0.03202
141216_at 6.407549 P 0.015164
141217_at 7.468318 P 0.001432
141218_at 6.544323 P 0.000491
141219_at 4.178143 P 0.002143

Total number of rows: 14010

Table truncated, full table size 409 Kbytes.




Supplementary file Size Download File type/resource
GSM95324.CEL.gz 2.5 Mb (ftp)(http) CEL
GSM95324.EXP.gz 310 b (ftp)(http) EXP

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