DPCR DUPLEXING METHOD IN A SINGLE FLUORESCENCE CHANNEL

Haoqing Zhang, Soňa Laššáková, Zhiqiang Yan, Xinlu Wang, Pavel Šenkyřík, Martina Gaňová, Honglong Chang, Marie Korabecna, Pavel Neuzil

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

Abstract

Here, we propose a novel single fluorescence channel-based dPCR duplexing method that combines a specific probe and intercalating dye to increase the difference in fluorescence amplitudes (F) between the two targets. We selected two targets, one detected only by the intercalating dye EvaGreen and the other detected by a combination of a FAM probe and EvaGreen. The results showed the difference in F values between two targets increased from 5% to 32%, when using the FAM probe compared with the detection of both amplicons using EvaGreen only. The added FAM probe enabled two-target discrimination using a single-color fluorescent channel.

Original languageEnglish
Title of host publicationMicroTAS 2022 - 26th International Conference on Miniaturized Systems for Chemistry and Life Sciences
PublisherChemical and Biological Microsystems Society
Pages1211-1212
Number of pages2
ISBN (Electronic)9781733419048
StatePublished - 2022
Event26th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2022 - Hybrid, Hangzhou, China
Duration: 23 Oct 202227 Oct 2022

Publication series

NameMicroTAS 2022 - 26th International Conference on Miniaturized Systems for Chemistry and Life Sciences

Conference

Conference26th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2022
Country/TerritoryChina
CityHybrid, Hangzhou
Period23/10/2227/10/22

Keywords

  • Digital PCR
  • single fluorescence channel duplexing

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