Materials and methods for quality-controlled two-color RT-QPCR diagnostic testing of formalin fixed embedded and/or fresh-frozen samples

Described herein are materials, methods, and kits enabling accurate and reproducible two-color reverse-transcription real-time quantitative PCR (RT-qPCR) for quality-controlled molecular diagnostic testing of samples that may contain degraded RNA. In certain aspects described herein are materials, methods, and kits for use in the molecular diagnostic testing of lung cancer in FFPE samples and/or fresh-frozen samples. Also described herein are materials and methods to control for inter-experimental variation occurring during two-color RT-qPCR amplification arising from variation in fluorescence specific activity, use of different thermocyclers, and inter-laboratory differences.
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Abstract/Description: Described herein are materials, methods, and kits enabling accurate and reproducible two-color reverse-transcription real-time quantitative PCR (RT-qPCR) for quality-controlled molecular diagnostic testing of samples that may contain degraded RNA. In certain aspects described herein are materials, methods, and kits for use in the molecular diagnostic testing of lung cancer in FFPE samples and/or fresh-frozen samples. Also described herein are materials and methods to control for inter-experimental variation occurring during two-color RT-qPCR amplification arising from variation in fluorescence specific activity, use of different thermocyclers, and inter-laboratory differences.
Subject(s): C12Q1/68
G06F19/24
Date Issued: 2016-12-27