This document describes the application of the Agilent 2100 bioanalyzer with the DNA 500 LabChip® kit for the development of meat speciation assays. By using the bioanalyzer to confirm the suitability of primer sets before probe production, the cost of probe production can be reduced.
This document details a method for developing real-time PCR assays for food authenticity, specifically for detecting and quantifying meat species. It highlights the use of the Agilent 2100 Bioanalyzer to verify that newly designed PCR primers function correctly and are specific before the costly production of fluorescent probes is undertaken. The authors demonstrate how this approach saves expenses by identifying non-specific primers early in the development process.
The kit is used to confirm that newly designed PCR primers are functioning correctly before the more expensive fluorescent probes are produced.
The PCR reactions were performed in 25-11 volumes using 300 nM of each primer.
The protocol consists of 30 cycles of a two-step reaction at 95 °C for 15 seconds and 60 °C for one minute, followed by a final 10-minute step at 72 °C.