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Sample GSM188732 Query DataSets for GSM188732
Status Public on Jun 06, 2007
Title GM07348
Sample type RNA
 
Source name lymphoblastoid cell line
Organism Homo sapiens
Characteristics Coriell cell line repository identifier: GM07348
http://locus.umdnj.edu/nigms/nigms_cgi/display.cgi?GM07348
CEPH sample
Gender: Female
Associated family: 1345-2
Family relationship: mother
Biomaterial provider Coriell Cell Repositories http://ccr.coriell.org/Sections/Search/Search.aspx?PgId=165&q=GM07348
Treatment protocol Untreated, baseline expression data.
Growth protocol Each sample was collected while in exponential growth.
Lymphoblastoid cell lines were centrifuged at 400 × g to remove media and 5 mL fresh lymphoblastoid cell media (LCL media) containing RPMI 1640 (Mediatech)/1% l-glutamine (Mediatech) plus 20% FBS (HyClone Laboratories) for the initial passage and then passaged every 48 h with LCL medium and 15% FBS. Cell suspensions were transferred to 25 cm2 flasks and incubated at 37oC in a 90% humidified 5% CO2 atmosphere. Cell lines were maintained at a concentration of 3.5-4.0×105 cells/mL and harvested following the 4th passage, only if viability >= 85%.
Extracted molecule total RNA
Extraction protocol Lymphoblastoid cell line suspensions were spun at 400 × g for 5 min to remove media. Cell pellets were washed twice with ice-cold PBS and stored at -80°C. Total RNA was extracted using RNeasy Plus Kits according to the manufacturers protocol.
Label biotin
Label protocol Ribosomal RNA was depleted from 1µg of total RNA using the RiboMinus Human/Mouse Transcriptome Isolation kit (Invitrogen Corp., Carlsbad, CA). cDNA was generated using the GeneChip WT cDNA Synthesis and Amplification Kit (Affymetrix, Inc., Santa Clara, CA) per manufacturer's instructions. cDNA was fragmented and end labeled using the GeneChip® WT Terminal Labeling Kit (Affymetrix, Inc.).
 
Hybridization protocol Approximately 5.5ug of labeled DNA target was hybridized to the Affymetrix GeneChip Human Exon 1.0 ST Array at 45oC for 16 hours per manufacturers recommendation (see http://www.affymetrix.com/products/arrays/exon_application.affx for additional information). Hybridized arrays were washed and stained on a GeneChip Fluidics Station 450.
Scan protocol All samples were scanned on a GCS3000 Scanner (Affymetrix, Inc.).
Description Gene expression on 176 HapMap cell lines (87 CEU and 89 YRI) was determined using the Affymetrix GeneChip Human Exon 1.0 ST Array.
Data processing dbSNP in Probe Filtering and Background Correction Protocol: The start and end coordinates of all probes of the Exon array were queried against the human genome (hg17). The coordinates for all SNPs were then queried in the dbSNP database (http://0-www-ncbi-nlm-nih-gov.brum.beds.ac.uk/projects/SNP/) (release 126) and then based on location, we identified probes harboring SNPs. In total, ~400,000 probes contained SNPs within their structure. Of the ~1.4 million probesets of the exon array, 255,676 contained at least one probe with SNP. Among these affected probesets, 105,000 harbored 2 or more probes with SNPs. Resulting probe signal intensity files were filtered by removing this group of 105,000probesets. After filtering, individual probe intensities were background corrected by subtracting the median intensity of a population of non-genomic probes with the same GC content to account for any non-specific hybridization. The resulting probe signal intensities were sketch quantile normalized over all 176 samples and gene-level expressions were summarized using the robust multi-array average (RMA) method with signals generated on a core set of exons. A constant of 16 was added to all probeset intensities for variance stabilization and summarized signals were then log2 transformed with a median polish.
 
Submission date May 14, 2007
Last update date Apr 26, 2009
Contact name Eileen Dolan
E-mail(s) edolan@medicine.bsd.uchicago.edu
Phone 773-702-4441
Fax 773-702-0963
Organization name University of Chicago
Department Medicine
Lab Dolan Lab
Street address 5841 S. Maryland
City Chicago
State/province IL
ZIP/Postal code 60657
Country USA
 
Platform ID GPL5175
Series (2)
GSE7761 Identification of Genetic Variants Contributing to Cisplatin-Induced Cytotoxicity using a Genome-wide Approach
GSE7851 Evaluation of Genetic Variation Contributing to Differences in Gene Expression between Populations
Relations
Alternative to GSM188914
Alternative to GSM245541

Data table header descriptions
ID_REF Transcript_probeset_id
VALUE Probe signal intensities were sketch quantile normalized over all 176 samples and gene-level expressions were summarized using the robust multi-array average (RMA) method with signals generated on a core set of exons. A constant of 16 was added to all probeset intensities for variance stabilization and summarized signals were then log2 transformed with a median polish.

Data table
ID_REF VALUE
3881686 9.103583032
2672712 8.17421047
2842570 8.759760682
3526544 6.56837493
2902531 5.726208167
2402942 6.382236771
3382216 6.678949955
3771800 10.16228577
2427469 5.063592578
2392945 7.333700913
2453006 5.720630811
2562821 7.910759705
3416651 7.040017798
3552083 5.903275724
3721851 8.085580565
2477438 5.066202544
2926969 4.736950595
3026969 7.233653313
3442176 7.99963382
3796335 8.111946332

Total number of rows: 17745

Table truncated, full table size 344 Kbytes.




Supplementary file Size Download File type/resource
GSM188732.CEL.gz 37.4 Mb (ftp)(http) CEL
Processed data included within Sample table

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