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11 April 2017 Droplet digital PCR for verification of interspersed refuge in midge tolerant wheat varietal blends
D.J. Perry, S.-J. Lee
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Abstract

Analysis of bulk ground samples by droplet digital PCR (ddPCR) was investigated as an alternative to individual kernel testing for assessment of interspersed refuge in midge [Sitodiplosis mosellana (Géhin)] tolerant wheat (Triticum aestivum L.) varietal blends. Four genotyping assays were selected such that at least one assay was informative for each of 15 varietal blends registered for production in Canada. The assays were examined in DNA of each of the constituent varieties to assess intravarietal polymorphism and in DNA mixtures simulating each varietal blend. Seed mixtures corresponding to a range of refuge proportions in two different varietal blends were prepared and assessed by ddPCR on two different platforms (RainDrop Digital PCR System, RainDance Technologies, Billerica, MA, and QX200 AutoDG Droplet Digital PCR System, Bio-Rad Laboratories, Hercules, CA). Both systems yielded refuge estimates that were very close to the targeted proportions across the range simulated. Standard deviations among estimates made with the QX200 were, on average, about 60% greater than those among corresponding estimates made with the RainDrop system, but in either case, the coefficients of variation generally remained under 5%. For a ddPCR-based assessment of interspersed refuge in midge tolerant wheat varietal blends, any advantage held by the RainDrop system with respect to precision may be offset by higher throughput achievable with the QX200.

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D.J. Perry and S.-J. Lee "Droplet digital PCR for verification of interspersed refuge in midge tolerant wheat varietal blends," Canadian Journal of Plant Science 97(2), 257-265, (11 April 2017). https://doi.org/10.1139/cjps-2016-0223
Received: 11 July 2016; Accepted: 1 September 2016; Published: 11 April 2017
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KEYWORDS
cécidomyie du blé
droplet digital PCR
interspersed refuge
mélanges variétaux
orange wheat blossom midge
PCR numérique par gouttelettes
refuge disséminé
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