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Yusuke Ii, Yuichi Uno, Michio Kanechi, and Noboru Inagaki

. To overcome this, a multiplex PCR reaction would be a useful way to amplify the target fragment together with a reaction control. Multiplex PCR has been developed for the detection of different pathogens ( Fraaije et al., 2001 ; Pastrik, 2000

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James J. Polashock and Nicholi Vorsa

DNA fingerprinting has been useful for genotypic classification of American cranberry (Vaccinium macrocarpon Ait.). Polymerase chain reaction (PCR) based methodologies including randomly amplified polymorphic DNA (RAPD) markers are relatively easy to use, and inexpensive as compared to other methods. However, RAPD markers have some limitations including seamless interlaboratory transferability and susceptibility to certain types of error. An alternative method, sequence characterized amplified regions (SCARs), was developed for cranberry germplasm analysis. Nine primer sets were designed from RAPD-identified polymorphic markers for use in two multiplex PCR reactions. These primer sets generated 38 markers across a cranberry germplasm collection. Estimates of genetic relatedness deduced from employment of the RAPD and SCAR methods were compared among 27 randomly chosen cranberry germplasm accessions. Although both methods produced comparable results above 0.90 coefficient of similarity, branches below this level exhibited variation in clustering. SCAR and RAPD markers can be employed for identifying closely related genotypes. However, the inferences of more distant genetic relationships are less certain. SCAR marker reactions provided more polymorphic markers on a per reaction basis than RAPD marker reactions and as such more readily separated closely related progeny. When SCAR primers were fluorescent dye-labeled for computerized detection and data collection, reduced marker intensity relative to unlabeled reactions was one problem encountered.

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Patrick D. O'Boyle, James D. Kelly, and William W. Kirk

100-bp ladder (Invitrogen). When multiplex PCR was used, SU91 (700 bp) and BC420 (900 bp) were distinguishable by size using ultraviolet light fluorescence. For multiplex PCR, the reaction conditions for the SAP6 SCAR marker was used ( Miklas et al

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Rose E. Palumbo, Wai-Foong Hong, Jinguo Hu, Charles Krause, James Locke, Richard Craig, David Tay, and Guo-Liang Wang

Pelargonium is one of the priority genera collected by the Ornamental Plant Germplasm Center (OPGC). In order to protect future breeders from a loss of genetic diversity, the OPGC collects heirloom cultivars, breeding lines, and wild species. The current Pelargonium collection consists primarily of cultivars originating from P. ×hortorum and P. ×domesticum. Our project was designed to analyze the current collection in order to facilitate the maintenance of a more-diverse core collection. We have expanded our TRAP (Target Region Amplified Polymorphism) analysis from 120 plants with one primer set to include 780 plants with four primer sets. Each primer set consists of a labeled arbitrary primer paired with a gene-specific primer, and two different fluorescent labels were used to allow multiplexed PCR reactions. We scored about 90 markers in each of the first two primer sets and about 60 markers in each of the second two. In comparisons between the phylogeny and the morphology and taxonomy of these plants, we show some matching clusters that may be explained by the breeding history of the plants.

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Chu-Hui Chiang, Tsong-Ann Yu, Shu-Fang Lo, Chao-Lin Kuo, Wen-Huang Peng, and Hsin-Sheng Tsay

the ITS sequences ( Baldwin et al., 1995 ; Hillis et al., 1991 ). Based on the ITS sequences, the multiplex PCR method can be used to amplify target-containing samples by mixing specific primers in a single PCR. This approach has been widely used to

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expedite sex determination in asparagus seedlings. Using a single step DNA extraction, excised roots of 19-day-old seedlings provided sufficient DNA for polymerase chain reaction (PCR) analysis without retarding seedling growth. Multiplex PCR using a

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Jason D. Zurn, Katie A. Carter, Melinda H. Yin, Margaret Worthington, John R. Clark, Chad E. Finn, and Nahla Bassil

identification and parentage confirmation. SSRs are abundant in plant genomes, cost-effective to use, amenable to automated scoring with software, and can be amplified in multiplex PCR ( Bassil et al., 2016 ; Peace, 2017 ; Weising et al., 2005 ). Moreover, SSRs

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Kate M. Evans, Bruce H. Barritt, Bonnie S. Konishi, Marc A. Dilley, Lisa J. Brutcher, and Cameron P. Peace

. Cova, V. Mott, D. Komjanc, M. Barbaro, E. Kodde, L. Rikkerink, E. Gessler, C. van de Weg, W.E. 2009 Development and test of 21 multiplex PCRs composed of SSRs spanning most of the apple

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Rafel Socias i Company, Àngel Fernández i Martí, Ossama Kodad, and José M. Alonso

the integration of self-compatible S -alleles from related species ( Gradziel et al., 2001 ). Screening efficiency and flexibility have also been greatly improved with the development of successful multiplex PCR techniques by Sánchez Pérez et al

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Tracie K. Matsumoto, Francis T.P. Zee, Jon Y. Suzuki, Savarni Tripathi, James Carr, and Bruce Mackey

in 55-1 transgene insertion. The multiplex PCR protocol to amplify the GUS (uidA) or PRSVcp within the transgene of 55-1 to indicate the transgenic event and papain as an internal control was used in this study ( Wall et al., 2004 ). The primer