Enzymatic labeling kit

Five ng of total RNA purified from each pool were amplified using Ovation Pico WTA System V2 (NuGEN Technologies, San Carlos, CA, USA) and labeled by Enzymatic Labeling Kit (Agilent Technologies). Three µg of purified-Cye3-labeled cDNA were hybridized to Human GE 4x44K v2 microarrays (Agilent Technologies) at 65 °C for 17 h. Slides were washed and scanned by Agilent G2505C scanner. Raw data were extracted using Feature Extraction (FE) software v10.7, GE1_1100_Jul11 protocol (Agilent Technologies).

An Agilent Enzymatic Labeling Kit was used according to the manufacturer’s instructions to label 0.5 μg of genomic DNA for each CGH array. Labeled DNA was hybridized to an Agilent-022060 SurePrint G3 Human CGH Microarray 4x180K (Agilent Technologies, Inc., Santa Clara, CA, USA); the Agilent Microarray Scanner and Feature Extraction v.10.7.3.1 (Agilent Technologies), were used according to manufacturer’s instruction to scan probed arrays. Control DNA was obtained from one Japanese individual who is an author (MU) of this study. We focused only on previously reported SCNAs of CDKN2A [14 (link)] and on CNVs of UGT2B17 that are associated with metabolism of nicotine [15 (link)]. The data described in this article have been deposited in NCBIs Gene Expression Omnibus (GEO) [16 ] and are accessible through GEO series accession number GSE47443.