TaqMan real-time PCR quantification strategy of CYP2D6 gene copy number for the LightCycler 2.0

TaqMan‐based real‐time polymerase chain reaction for detection of FUT2 copy number variations: identification of novel Alu‐mediated deletion

“…We performed real‐time PCR to amplify the following three regions simultaneously: 1) the promoter region of the FUT2 allele (Prom) that is absent in the se fus allele but present in se del and se del2 alleles; 2) the 5′‐flanking region of the FUT2 coding region (FUT2‐5′), which lacks the se fus , se del , and se del2 alleles; and 3) Exon 12 of the ALB (which is assumed to always have two copies/genome) as the internal control to compare the intensity of the amplification signals 19 . The relative positions of Prom, FUT2‐5′, and the deleted regions of three recombination alleles of the FUT2 gene are illustrated in Fig.…”

Single tube genotyping of CYP2A6 gene deletion based on copy number determination by quantitative real-time PCR

“…By using this technical strategy, assays designed for the determination of the gene deletion or duplication of CYP2D6 has been successfully developed and validated (Nguyen et al, 2009;Schaeffeler et al, 2003). However, the complex genetic architecture complementary to its significant homology with CYP2A7 and CYP2A13 challenges the application of this strategy in the detection of CYP2A6 gene deletions.…”

“…Commercial assays using quantitative PCR and microarray for CYP2A6 gene copy number determination are reported to work well (Martis et al, 2013), but such approaches the price prohibitive and the oligonucleotide sequences of these assays have not been published. Quantitative real-time PCR has been successfully applied in gene copy number determination in previous studies (Aarskog and Vedeler, 2000;Nguyen et al, 2009;Schaeffeler et al, 2003). In this work, we designed and validated a sensitive and specific assay for genotyping CYP2A6 gene deletion that was based on gene copy number determination by quantitative real-time PCR.…”

Identification of Novel Copy Number Variations of VCAN Gene in Three Chinese Families with Wagner Disease

“…For each sample, at least three PCR reactions were performed to ensure reproducibility. The differences in expression between patients and controls were calculated by using the 2 ΔΔCp method [ 17 ]. Student’s t -test was used for the statistical analysis of the qPCR data by GraphPad (GraphPad Software, San Diego, CA, USA).…”