Sureprint g3 human cgh microarray 8 60k chips

DNA quality was checked, and then the qualified DNA samples were detected using SurePrint G3 Human CGH Microarray 8 × 60k chips (Agilent, Santa Clara, USA) according to the operating instruction. The microarrays were scanned and analyzed by microarray scanner (Agilent, Santa Clara, USA) and other supporting software. Subsequently, quantitative polymerase chain reaction (qPCR) was used to validate these candidate CNVs using StepOne-type fluorescent qPCR instrument (ABI, Vernon, USA). The primers used in qPCR are shown in Table 1. Microarray results were further compared with UCSC, DECIPHER, DGV, ISCA, and OMIM databases to identify pathogenicity of CNVs.

DNA quality was checked and the qualified DNA samples were then detected using SurePrint G3 Human CGH Microarray 8 × 60 K chips (Agilent Technologies Inc., Santa Clara, CA, USA). After lysis, labeling Cy‐dUTP and Cy‐dUTP, purification, hybridization and washing, scanning and data extraction were performed using a Microarray scanner (Agilent Technologies Inc.) and relevant software that could indicate CNVs with three consecutive probe log₂ values greater than 0.25 or less than −0.25. Most of pathogenic CNVs (99.34%) are larger than 300 kb.9 CNVs greater than 200 kb are generally detected.10 In the present study, CNVs with chromosomal aneuploidy and greater than 200 kb were detected, and so the possibility of minor anomalies occurring in chromosome structures or gene fragment was not excluded. Chip sequence information was from hg19. The microarray results were further compared with the University of California Santa Cruz (UCSC), Database of Chromosomal Imbalance and Phenotype in Humans using Ensembl Resources (DECIPHER), Database of Genomic Variants (DGV), Institute of Singapore Chartered Accountants (ISCA) and Online Mendelian Inheritance in Man (OMIM) databases to analyze the pathogenicity of CNVs.