BioPrime™ Array CGH Genomic Labeling Module

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		Label all samples at one time to avoid day-to-day variations. The labeled samples can be stored at -80°C upto 3 months and successive hybridizations

Downstream tips
Label all samples at one time to avoid day-to-day variations. The labeled samples can be stored at -80°C upto 3 months and successive hybridizations

Publication protocol

CGH array chips
For the CGH array experiments, 1 μg of purified patient DNA and control DNA were digested with 10U Alui and 10U RsaI and differentially labeled with cyanine-5 (Cy5) and cyanine-3 (Cy3) fluorescent dyes (PerkinElmer, Waltham, Massachusetts, USA) by random priming with the Bioprime CGH Array genomic labeling module (Invitrogen, Carlsbad, California, USA). Hybridization was performed for at least 20 h at 65°C in a rotating microarray hybridization chamber and then samples were washed according to the manufacturer's protocols (Agilent Technologies, Santa Clara, California, USA). The slide hybridization results were scanned into image files using a GenePix 4000B microarray scanner (Molecular Devices, Sunnyvale, California, USA). Then, Agilent's Feature Extraction v9.1 was used to quantify the array features, and the CGH analytics software was used to analyze the text file outputs for relative copy number changes. Common copy number variations were assessed by comparing these sample results with CNV databases and parental CGH patterns, when available.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for BioPrime™ Array CGH Genomic Labeling Module below.

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