The use of standards in quantitative PCR (qPCR) is essential, especially in forensic cases, to determine the concentration of DNA in an unknown sample. The standard curve for Quantifiler Trio is commonly made up of a ten-fold serial dilution of the known DNA standard in the qPCR kit. Due to the known concentration of the standard and the serial dilution, the threshold cycle values of the standards can be compared to the cycle threshold values of the unknown samples to determine their concentrations. Use of the Quantifiler Trio kit provides the concentration of total human DNA, including both small autosomal and large autosomal amounts which are used to calculate a degradation ratio, and also determines the concentration of male DNA in the sample. Each aspect has its own standard curve to determine the DNA concentration.
However, serial dilutions cause variance between runs, even when using the same samples, due to small pipetting differences or errors which can make sample reruns and verifying data difficult. Creating a virtual curve by using data from multiple serial dilutions would minimize or eliminate the variance between runs. This is accomplished by taking the averages of slopes and y-intercepts of the standards to creature the virtual curve.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/44012 |
| Date | 10 March 2022 |
| Creators | Lively, Brianna |
| Contributors | Cotton, Robin W. |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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