SurePrint G3 Human CGH Microarray Kit, 4x180K

Protocol tips

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Avoid touching the surface of the glass with your fingers. Skin oils and other substances, such as lotions or ink, can fluoresce. If the surface of the glass is noticeably dirty, it can be carefully cleaned with a nonabrasive laboratory tissue.

Upstream tips
Avoid touching the surface of the glass with your fingers. Skin oils and other substances, such as lotions or ink, can fluoresce. If the surface of the glass is noticeably dirty, it can be carefully cleaned with a nonabrasive laboratory tissue.

Publication protocol

In total, 25 patients diagnosed with RP, STGD1 and CORD and 2 unaffected family members, were analyzed for copy number variations (CNVs) using comparative genomic hybridization arrays (aCGH) as previously described (Birnbach et al., 1994). In brief, aCGH was performed using standard protocols with SurePrint G3 Human CGH 4x180K microarray platform (Agilent Technologies, Santa Clara, CA, USA). Each array contains approximately 180,000 60-mer oligonucleotide probes. Labeling was performed using the SureTag DNA labeling kit (Agilent technologies). The microarrays were hybridized in a rotisserie oven at 67 °C for 24 h according to manufacturer's recommendations. The arrays were scanned using an Agilent DNA microarray scanner (G2565CA, software version 4.8.4.1) and the spot intensities were measured using the Agilent Feature Extraction Software (version 10.7.3.1). Raw data were analyzed by Agilent Cytogenomics software (version 3.0.2.11). The cutoff for recording an alteration was ≥ 4 consecutive probes. For the interpretation of the results, the following databases were used: database of genomic variants (http://projects.tcag.ca/variation/), DECIPHER database (http://www.sanger.ac.uk/PostGenomics/decipher/), and the International Standards for Cytogenomic Arrays (https://www.iscaconsortium.org/).

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Manufacturer protocol

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