The array CGH was performed using an Agilent Human Genome CGH microarray 180 K kit (Agilent Technologies Inc. Shanghai, China) with a resolution of 6.4 kb. For each sample, 1.5 μg of genomic DNA was used for each array CGH experiment. The array CGH experiments were performed according to the manufacturer’s instruction as described previously
[12 (link)]. Briefly, genomic DNA from patients and controls was digested with the restriction enzymes AluI and RsaI (Promega) and fluorescently labeled with Cy5 (patients) or Cy3 (controls) using the Agilent DNA Labeling Kit. Labeled DNAs were combined, denatured, pre-annealed with Cot- DNA (Invitrogen) and blocking reagent (Agilent), and then hybridized to the arrays for 40 hours in a rotating oven (Agilent Technologies) at 65°C and a speed of 20 rpm. After performing the hybridization step and the recommended washes, the arrays were scanned at 5 μm resolution with an Agilent G2505A scanner. The images were analyzed using the Feature Extraction software (Agilent Technologies, Shanghai, China). All array data that passed the quality metrics were further processed by the DNA Analytics software.
[12 (link)]. Briefly, genomic DNA from patients and controls was digested with the restriction enzymes AluI and RsaI (Promega) and fluorescently labeled with Cy5 (patients) or Cy3 (controls) using the Agilent DNA Labeling Kit. Labeled DNAs were combined, denatured, pre-annealed with Cot- DNA (Invitrogen) and blocking reagent (Agilent), and then hybridized to the arrays for 40 hours in a rotating oven (Agilent Technologies) at 65°C and a speed of 20 rpm. After performing the hybridization step and the recommended washes, the arrays were scanned at 5 μm resolution with an Agilent G2505A scanner. The images were analyzed using the Feature Extraction software (Agilent Technologies, Shanghai, China). All array data that passed the quality metrics were further processed by the DNA Analytics software.
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