|
| Status |
Public on Jun 24, 2010 |
| Title |
TAX577565 |
| Sample type |
genomic |
| |
|
| Channel 1 |
| Source name |
Breast tumor
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Breast tumor
hu subtype: Normal
familial status: familial
er: er_pos
osbin: 0
os: 10.84109589
pgr: pgr_neg
grade: NA
genomic subtype: amplifier
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Genomic DNA from fresh frozen breast tumor tissue was extracted using overnight digestion with proteinase K at 55C, followed by repeated phenol-chloroform purification steps. DNA concentrations were measured using a Nanodrop.
|
| Label |
Cy3
|
| Label protocol |
Fluorescently labeled DNA targets for hybridization were prepared according to manufacturers' instructions using the Invitrogen Bioprime aCGH Genomic Labeling Module as described (PMID:17334996).
|
| |
|
| Channel 2 |
| Source name |
promega male reference
|
| Organism |
Homo sapiens |
| Characteristics |
reference: promega male reference
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Genomic DNA from fresh frozen breast tumor tissue was extracted using overnight digestion with proteinase K at 55C, followed by repeated phenol-chloroform purification steps. DNA concentrations were measured using a Nanodrop.
|
| Label |
Cy5
|
| Label protocol |
Fluorescently labeled DNA targets for hybridization were prepared according to manufacturers' instructions using the Invitrogen Bioprime aCGH Genomic Labeling Module as described (PMID:17334996).
|
| |
|
| |
| Hybridization protocol |
BAC aCGH microarrays were hybridized as described (PMID: 17334996)
|
| Scan protocol |
Arrays were washed as described (PMID: 17334996) and fluorescence was recorded using an Agilent G2565AA microarray scanner (Agilent Technologies).
|
| Data processing |
TIFF images were analyzed using the Gene Pix Pro software (Axon Instruments, Foster City, CA), and the quantified data matrix was loaded into a local installation of BioArray Software Environment (BASE) (http://base.thep.lu.se). Quantified intensities for tumor sample were stored as channel 1 (ch1) whereas quantified intensities for reference sample were stored as channel 2 (ch2). Median spot pixel values were used to calculate background corrected intensities (Int1 and Int2). Spots were removed and regarded as missing values if either: (1) flagged during image-analysis, (2) background corrected intensity value >64999 signal units, (3) background corrected intensity value <0 signal units, or (4) Signal to noise ratio <5 in either Int1 or Int2 channel. For spots, log ratio (M) was calculated as log2(Int1/Int2) and average intensity (A) was calculated as log10(Int1*Int2)/2. Normalization was performed by applying popLowess (PMID: 17953745). Spots from dye-swap hybridization were merged using geometric mean of ratios after normalization.
|
| |
|
| Submission date |
Jun 06, 2010 |
| Last update date |
Jun 24, 2010 |
| Contact name |
Johan Staaf |
| Organization name |
SCIBLU - Swegene Centre for Integrative Biology at Lund University
|
| Street address |
Medicon Village
|
| City |
Lund |
| ZIP/Postal code |
SE-223 81 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL4723 |
| Series (2) |
| GSE22133 |
Genomic Subtypes of Breast Cancer Identified by Array Comparative Genomic Hybridization |
| GSE32578 |
Characterization of amplification patterns and target genes at chromosome 11q13 in CCND1-amplified sporadic and familial breast tumors |
|
