The aCGH platform from Agilent Technologies was employed in this study. Custom-designed 44 K as described previously [33 (link)] and ISCA designed 180 K oligonucleotide microarrays were used. Both designs had similar genomic coverage for interrogation of over 100 genetic disorders but were different in probe density giving average genome-wide resolution of 100 kb and 25 kb, respectively.
Experimental procedures were performed according to manufacturer’s description. Briefly, one microgram of patient’s DNA and normal female control DNA (Promega) were differentially labeled with Cy5 and Cy3 respectively using Agilent SureTag Complete DNA Labeling Kit (Agilent Technologies, USA). Labeled DNA was then cleaned by purification columns (Agilent Technologies, USA) and hybridized on microarray for 24 hours. Microarray washing and scanning was performed using Agilent Oligo aCGH Wash Buffers (Agilent Technologies, USA) and Agilent Microarray Scanner (Agilent Technologies, USA) according to manufacturer’s instructions. All pathogenic CNVs were further validated on NimbleGen CGX-135 K array which were designed by Signature Genomics (Perkin Elmer, USA) following manufacturer’s instructions.
Experimental procedures were performed according to manufacturer’s description. Briefly, one microgram of patient’s DNA and normal female control DNA (Promega) were differentially labeled with Cy5 and Cy3 respectively using Agilent SureTag Complete DNA Labeling Kit (Agilent Technologies, USA). Labeled DNA was then cleaned by purification columns (Agilent Technologies, USA) and hybridized on microarray for 24 hours. Microarray washing and scanning was performed using Agilent Oligo aCGH Wash Buffers (Agilent Technologies, USA) and Agilent Microarray Scanner (Agilent Technologies, USA) according to manufacturer’s instructions. All pathogenic CNVs were further validated on NimbleGen CGX-135 K array which were designed by Signature Genomics (Perkin Elmer, USA) following manufacturer’s instructions.