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The
Functional Genomics Core Laboratory offers "full service"
microarray studies using spotted cDNA, spotted oligonucleotide,
and antibody microarray platforms. The Microarray Services Unit
develops and maintains numerous microarray-associated protocols
encompassing all of the technical aspects of microarray experiments.
Raw data are obtained by scanning hybridized arrays with a Packard
ScanArray 5000XL fluorescent scanner.
The
spotted cDNA microarrays consist of full-length gene sequences
and ESTs derived from the NIA mouse cDNA cloneset and the Research
Genetics human library. The cDNA-based arrays contain between
9,000 to 15,000 mouse, rat, or human gene sequences. The cDNA
microarrays are supplied by the Microarray Centre at the Ontario
Cancer Institute
Spotted
oligonucleotide microarrays provide the advantage of greater specificity
and improved quantitative accuracy over spotted cDNA microarrays.
Oligonucleotide-based arrays consist of between 1,000 and 20,000
mouse, rat, or human gene sequences. The oligonucleotide-based
microarrays are obtained from commercial vendors such as Mergen
LTD., Amersham Biosciences (CodeLink), and Agilent.
Antibody
arrays are obtained from BD Biosciences/Clontech (Palo Alto, CA).
These microarrays contain 380 distinct monoclonal antibodies that
are covalently immobilized on a glass slide.
Specific
microarray-related services include, but are not limited to:
- Consultation
on experimental design and microarray platform selection
- Quantitative
RT-PCR (SYBR Green or TaqMan) for pre-array evaluation of samples
- Total
RNA extraction
- RNA
amplification (aRNA)
- Quality
evaluation of total RNA and aRNA
- Cy3/Cy5
(or other dyes) nucleic acid labeling
- Protein
extraction and Cy3/Cy5 protein labeling
- Microarray
hybridization
- Microarray
scanning
- Quantitative
RT-PCR (TaqMan) for validation of microarray results
- Assistance
on grant or protocol submission (e.g., preparing microarray
study budget and providing protocol descriptions)
- Manuscript
review and preparation
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