qBiomarker Copy Number PCR Assays
For locus-specific analysis of copy number variations and alterations
qBiomarker Copy Number PCR Assays provide a method for obtaining specific, accurate, reproducible, and easy-to-interpret copy number change results for an individual gene or region of interest (GOI or ROI). Every assay has been bench-verified and is ready-to-use in microarray follow-up studies, specific target screening, and related studies.
All qBiomarker Copy Number PCR Assays are wet bench-tested for several characteristics affecting the accuracy of real-time PCR results: specificity, wide dynamic range, and uniformly high amplification efficiency. Laboratory verification of assay quality ensures that qBiomarker Copy Number PCR Assays deliver reliable results.
qBiomarker Copy Number PCR Assays accurately identified aneuploidy in cell lines containing chromosomal aberrations previously identified by cytogenetic methods. Assays for AR and MECP2, which are both on the X chromosome, correctly quantified gene copy number in cell lines with 1, 3, and 4 copies of the X chromosome.
All qBiomarker Copy Number PCR Assays are designed in unique regions of the genome. The qBiomarker Multicopy Reference Copy Number PCR Assay (MRef), which can be purchased separately, provides superior normalization and is recommended for use with the qBiomarker Copy Number PCR Assays. The MRef assay recognizes a stable sequence that appears in the human genome over 40 times, and whose copy number is not affected or minimally affected by local genomic changes. Inclusion of this reference assay during testing allows use of the ΔΔCT method to accurately make copy number calls or relative copy number change calls for specific targets. The all-in-one, ready-to-use qBiomarker SYBR® Green PCR Mastermixes are optimized to provide maximum locus-specific amplification with minimal primer dimers.
Isolate genomic DNA from fresh, frozen, or FFPE samples. Combine an aliquot of each sample with the appropriate ready-to-use qBiomarker SYBR Green Mastermix and qBiomarker Copy Number PCR Assay. In another tube, mix an aliquot of each sample with the mastermix and the qBiomarker Multicopy Reference Copy Number PCR Assay. Run in a real-time cycler and determine copy number changes by the ΔΔCT method by comparing the test sample with a reference genome using the complimentary data analysis tool.
qBiomarker Copy Number PCR Assays are highly suited for accurate detection of copy number alterations or variations at individual loci using fresh, frozen, or FFPE samples.
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