Ovation® WGA System
Ovation WGA System offers a robust and simple amplification technology based on SPIA® (Single Primer Isothermal Amplification). This amplification system faithfully replicates genomic DNA from clinical samples, providing a valuable resource for a variety of downstream applications including copy number change analysis using real-time quantitative PCR (qPCR) or microarrays. Because of the simplicity and robustness of the procedure, accurate and quantitative results are obtained with minimal sample or allele dropouts and without the need for customization or optimization.
Starting with 10 ng genomic DNA, microgram quantities of amplified SPIA product can be prepared in approximately four hours. The Ovation WGA System (Part No. 6100) provides optimized reagent mixes to process 12 DNA samples.
Consistent Performance out of the Box: 87 qPCR markers were designed to be evenly spaced across the human genome. Replicate amplification was performed starting with 10 ng of genomic DNA isolated from cultured cells. No allele or sample dropout was observed in the first experiment in the absence of any customization or optimization. Average Ct values from TaqMan assays were plotted, demonstrating highly reproducible results from two independent amplification reactions.
A Broad Range of Downstream Quantitative Applications: The linear nature of SPIA technology allows accurate quantification and is particularly well suited for copy number analysis. Genomic DNA was isolated from six Trisomy 21 patient and six control blood samples. TaqMan assays were carried out with qPCR markers from a Chromosome 21 locus and a control Chromosome 12 locus (GAPDH). The expected value of ΔCt between the two markers from the trisomy 3:2 chromosomal imbalance should be 0.6. ΔCt values of 0.6 were consistently obtained from the SPIA-amplified and unamplified materials.
Accurate Gain and Loss Analysis Results with aCGH: Another powerful whole genome copy number analysis technology is Array Comparative Genome Hybridization (aCGH) for surveying the entire genome at high resolution. Comparable detection of copy number abnormalities was obtained from the amplified or unamplified samples.