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Academic literature on the topic 'KBioscience's Allele-Specific PCR (KASP)'
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Journal articles on the topic "KBioscience's Allele-Specific PCR (KASP)"
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Majeed, Uzma, Essam Darwish, Shoaib Ur Rehman, and Xueyong Zhang. "Kompetitive Allele Specific PCR (KASP): A Singleplex Genotyping Platform and Its Application." Journal of Agricultural Science 11, no. 1 (2018): 11. http://dx.doi.org/10.5539/jas.v11n1p11.
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Single nucleotide polymorphism (SNP) can be detected by mining sequence databases or by using different singleplex or multiplex SNP genotyping platforms. Development of high-throughput genotyping molecular markers can be instrumental towards maximizing genetic gain. In this review we provide an overview of Kompetitive Allele Specific PCR (KASP) genotyping platform requirements and its application that might be helpful in KASP marker development. This literature further illustrates the possibilities to design KASP primers. Several research institutes routinely using KASP platform, producing in excess of humungous data points yearly for breeding cultivars and as well as for medical and commercial purposes. KASP genotyping technology offers cost effectiveness and high throughput molecular marker development platform. Conventional molecular markers can be converted into more robust and high throughput KASP markers. More than 2000 published references clearly show the popularity of KASP technology among the researchers.
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Cheon, Kyeong-Seong, Young-Min Jeong, Hyoja Oh, et al. "Development of 454 New Kompetitive Allele-Specific PCR (KASP) Markers for Temperate japonica Rice Varieties." Plants 9, no. 11 (2020): 1531. http://dx.doi.org/10.3390/plants9111531.
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Temperate japonica rice varieties exhibit wide variation in the phenotypes of several important agronomic traits, including disease resistance, pre-harvest sprouting resistance, plant architecture, and grain quality, indicating the presence of genes contributing to favorable agronomic traits. However, gene mapping and molecular breeding has been hampered as a result of the low genetic diversity among cultivars and scarcity of polymorphic DNA markers. Single nucleotide polymorphism (SNP)-based kompetitive allele-specific PCR (KASP) markers allow high-throughput genotyping for marker-assisted selection and quantitative trait loci (QTL) mapping within closely related populations. Previously, we identified 740,566 SNPs and developed 771 KASP markers for Korean temperate japonica rice varieties. However, additional markers were needed to provide sufficient genome coverage to support breeding programs. In this study, the 740,566 SNPs were categorized according to their predicted impacts on gene function. The high-impact, moderate-impact, modifier, and low-impact groups contained 703 (0.1%), 20,179 (2.7%), 699,866 (94.5%), and 19,818 (2.7%) SNPs, respectively. A subset of 357 SNPs from the high-impact group was selected for initial KASP marker development, resulting in 283 polymorphic KASP markers. After incorporation of the 283 markers with the 771 existing markers in a physical map, additional markers were developed to fill genomic regions with large gaps between markers, and 171 polymorphic KASP markers were successfully developed from 284 SNPs. Overall, a set of 1225 KASP markers was produced. The markers were evenly distributed across the rice genome, with average marker density of 3.3 KASP markers per Mbp. The 1225 KASP markers will facilitate QTL/gene mapping and marker-assisted selection in temperate japonica rice breeding programs.
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Chang, Ching-Chi, Benji Brayan I. Silva, Huai-Ying Huang, et al. "Development and Validation of KASP Assays for the Genotyping of Racing Performance-Associated Single Nucleotide Polymorphisms in Pigeons." Genes 12, no. 9 (2021): 1383. http://dx.doi.org/10.3390/genes12091383.
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Pigeon racing’s recent upturn in popularity can be attributed in part to the huge prize money involved in these competitions. As such, methods to select pigeons with desirable genetic characteristics for racing or for selective breeding have also been gaining more interest. Polymerase chain reaction—restriction fragment length polymorphism (PCR-RFLP) for genotyping-specific genes is one of the most commonly used molecular techniques, which can be costly, laborious and time consuming. The present study reports the development of an alternative genotyping method that employs Kompetitive Allele Specific Polymerase Chain Reaction (KASP) technology with specifically designed primers to detect previously reported racing performance-associated polymorphisms within the LDHA, MTYCB, and DRD4 genes. To validate, KASP assays and PCR-RFLP assays results from 107 samples genotyped for each of the genes were compared and the results showed perfect (100%) agreement of both methods. The developed KASP assays present an alternative rapid, reliable, and cost-effective method to identify polymorphisms in pigeons.
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Yonet, Nilay, Yıldız Aydin, Goksel Evci, and Ahu Altinkut Uncuoglu. "Genomic Evaluation of Sunflower Broomrape (Orobanche Cumana) Germplasm by KASP Assay." Helia 41, no. 68 (2018): 57–72. http://dx.doi.org/10.1515/helia-2017-0016.
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AbstractOrobanche cumana Wallr. is a holoparasitic plant for only sunflower, hence it is called as sunflower broomrape. Yield loss created by O. cumana which is generally 50 % can reach to 100 %. In this study, it was planned to perform molecular characterization of O. cumana germplasm as nine locations of Thrace region obtained from Trakya Agricultural Research Institute by using Single Nucleotide Polymorphism (SNP) markers, widely used in plant breeding programs, in Competitive Allele Specific PCR (KASP) assay which is a fluorescent tagged allele specific PCR method based, economic, reliable and easily repeatable genotyping technology. Databases and literature were scanned to spot variations on O. cumana genome which is not known clearly. So far, four SSR (Simple Sequence Repeat) marker (Ocum-197, Ocum-006, Ocum-023 and Ocum-151) regions showing polymorphic pattern were used for searching possible SNPs. Primer pairs were designed for amplification of the regions possibly having SNPs and PCR amplifications with these primer pairs were performed and 1 candidate deletion was detected on the amplicon which was amplified by Ocum-197 SSR marker. Following, the deletion was converted to KASP primers and KASP assay was performed. The deletion marker, Del-197, has grouped the samples from nine locations in the resulting allelic discrimination plot and infestation was performed according to this grouping, As a conclusion, Del-197 is considered as a selective marker for the ability to rapidly assay allelic variation at DNA markers for O. cumana populations that have effects on infestation results were evaluated as races, F, G, H and I in Thrace region.
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Yuan, Jiazheng, Zixiang Wen, Cuihua Gu, and Dechun Wang. "Introduction of High Throughput and Cost Effective SNP Genotyping Platforms in Soybean." Plant Genetics, Genomics, and Biotechnology 2, no. 1 (2017): 90–94. http://dx.doi.org/10.5147/pggb.v2i1.155.
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We presented here the application of two in-plate SNP (single nucleotide polymorphism) genotyping platforms for soybean plants [Glycine max (L.) Merr.], KASP® (Kompetitive Allele Specific PCR genotyping, LGC Genomics) and TaqMan® (Life Technologies) respectively. These two systems offer us an ability to determine the genotypes of 384 individual samples accurately and efficiently by allele specific PCR in a single plate using typical PCR conditions. Both of the systems require small quantity of genomic DNA obtained from a simple DNA extraction. The genomic sequences containing target SNPs can easily be used as a basic blueprint to design the probes and primers of KASP® and TaqMan® assays whether the sequences are obtained from the genome sequence of soybean William 82 (Wm82.a2.v1), Illumina Soy50k SNPs, or parallel resequencing. Moreover, we listed the pros and cons of the two systems and explained the principles behind the platforms. The high call rate and clear clustering separation of the SNPs can be readily obtained from these platforms without conducting any assay optimization processes. These platforms can routinely be performed on 96/384-well plate format with or without an automation procedure. Therefore, these platforms are especially suitable for the SNP genotyping on a particular trait with a large sample size, gene fine mapping, and marker assisted selection. Further, they require little hands-on experience and achieve per-site and per-individual costs below that of current SSR, AFLP, RFLP, and SNP chip technologies. The platforms can be used for genotyping on a wide range of organisms due to their simplicity and flexibility of handling. Meanwhile, we also especially presented some of the advantages using KASP® SNP genotyping pipeline, which was cost effective in the selection of allele specific assay and therefore, efficiently facilitated the soybean genotyping across large numbers (thousands or more) of individual lines for a great range of markers (hundreds to thousands) in our laboratory.
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Sejake, Thulo, Nemera Shargie, Riann Christian, Assefa B. Amelework, and Toi J. Tsilo. "Genetic diversity in sorghum (Sorghum bicolor L. Moench) accessions using SNP based Kompetitive allele-specific (KASP) markers." June 2021, no. 15(06):2021 (June 10, 2021): 890–98. http://dx.doi.org/10.21475/ajcs.21.15.06.p3088.
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Genetic diversity analysis is an important component in conventional and marker-assisted breeding. The objective of this study was to assess the level of genetic diversity among 100 sorghum accessions, which were selected randomly from the Sorghum National Germplasm Bank maintained at Agricultural Research Council, South Africa. A total of 136 Kompetitive Allele-Specific PCR (KASP) markers were used in this study. The KASP markers were previously derived from single-nucleotide polymorphic (SNP) analysis of the world-wide sorghum accessions by other research groups. A total of 110 KASP markers were polymorphic and recorded an average polymorphic information content (PIC) value of 0.3, which indicated high level of discrimination of the markers. The markers had an average gene diversity and observed heterozygosity of 0.3 and 0.10, respectively. Analysis of molecular variance revealed a significantly high variation among accessions (83% and 89%) than within accessions (10% and 11%) based on breeding status and geographic origin, respectively. Genetic distance varied from 0.0 between SA0672 and SA0673, SA1282 and SA0670 to 0.57 between Hakika and SA1442 with an average mean of 0.30. The dendrogram and model-based population analysis identified three and four distinct groups in 95 sorghum accessions, respectively. These results imply the presence of genetic diversity and lack of genetic bottleneck within the National Sorghum Germplasm Bank, which could be highly relevant for sorghum breeding and germplasm maintenance
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Anuarbek, Shynar, Saule Abugalieva, and Yerlan Turuspekov. "Validation of Bread Wheat KASP Markers in Durum Wheat Lines in Kazakhstan." Proceedings of the Latvian Academy of Sciences. Section B. Natural, Exact, and Applied Sciences. 73, no. 5 (2019): 462–65. http://dx.doi.org/10.2478/prolas-2019-0071.
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Abstract Development of efficient DNA markers plays an important role in modern breeding projects of many crops, including cultivated hexaploid bread wheat (BW) and tetraploid durum wheat (DW). Findings of genome-wide association studies on major polyploid crops, such as BW, may also help in molecular breeding studies in relative cultivated species with a similar genetic background, including DW. Therefore, the validation of identified quantitative trait loci or marker-trait associations is an important preliminary step in marker-assisted selection (MAS) oriented projects. In this study, thirty-two SNP (single nucleotide polymorphism) markers of six agronomic traits identified in BW, harvested in Kazakhstan, were converted to KASP (Kompetitive Allele-Specific PCR) as-says. Generated 32 KASP assays were used in the analysis of 29 DW accessions from Kazakhstan. Firstly, the group of DW accessions was tested using replicated and randomised one-metre blocks in field conditions of southeast Kazakhstan and evaluated for main agronomic traits. The analysis showed that 14 KASP assays were polymorphic in the scoring of 29 DW accessions. The t-test suggested that the segregation in eight KASP assays was significantly associated with five agronomic traits. The study confirms robustness of KASP assays in MAS of DW breeding projects for the improvement of yield potential.
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Brabham, Chad, Jason K. Norsworthy, and Fidel González-Torralva. "Presence of the HPPD Inhibitor Sensitive 1 Gene and ALSS653N Mutation in Weedy Oryza sativa Sensitive to Benzobicyclon." Plants 9, no. 11 (2020): 1576. http://dx.doi.org/10.3390/plants9111576.
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Benzobicyclon has shown varying results in controlling weedy rice, including those with imidazolinone (IMI) resistance. Tolerance to benzobicyclon in cultivated japonica rice, but not indica or aus-like cultivars, is conferred by a fully functional HPPD Inhibitor Sensitive 1 (HIS1) gene. Herein, a diagnostic Kompetitive Allele Specific PCR (KASP) assay was developed to predict the HIS1 genotype of weedy rice plants from 37 accessions and correlated to their response to benzobicyclon in the field. Two-thirds of the 693 weedy rice plants screened were tolerant to benzobicyclon (371 g ai ha−1, SC formulation) at 30 days after treatment (DAT). Thirty-four percent of plants were homozygous for the HIS1 allele and 98% of these plants exhibited field tolerance. However, the his1 genotype did not always correlate with field data. Only 52% of his1 plants were considered sensitive, indicating that the single nucleotide polymorphisms (SNPs) chosen in the KASP assay are not a reliable tool in predicting his1 homozygous plants. In an additional experiment, 86% of the 344 plants with at least one copy of the ALSS653N trait harbored a HIS1 allele, suggesting fields infested with IMI herbicide-resistant weedy rice are unlikely to be controlled with benzobicyclon.
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Jang, Woojong, Hyun Oh Lee, Jang-Uk Kim, et al. "Complete Mitochondrial Genome and a Set of 10 Novel Kompetitive Allele-Specific PCR Markers in Ginseng (Panax ginseng C. A. Mey.)." Agronomy 10, no. 12 (2020): 1868. http://dx.doi.org/10.3390/agronomy10121868.
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Panax ginseng C. A. Mey., a perennial herb belonging to the family Araliaceae, is a valuable medicinal plant with distinctive biological characteristics. However, comprehensive analyses of the mitochondrial genome (mitogenome) are lacking. In this study, we sequenced the complete mitogenome of ginseng based on long-read data from the Nanopore sequencing platform. The mitogenome was assembled into a “master circle” form of 464,705 bp and contained 72 unique genes. The genome had three large repeat regions, and 10.42% of the sequences were mitogenome sequences of plastid origin (MTPTs). In total, 278 variants (213 SNPs and 65 InDels) were discovered, most of which were identified in intergenic regions. The MTPT regions were mutational hotspots, harboring 74.5% of the variants. The ginseng mitogenome showed a higher mutation rate than that of the chloroplast genome, and this pattern is uncommon in plants. In addition, 10 Kompetitive allele-specific PCR (KASP) markers were developed from 10 SNPs, excluding those in MTPT regions. These markers accurately identified the genotypes of 59 Korean ginseng accessions and elucidated mitogenome diversity. These results provide insight into organellar genomes and genetic diversity in ginseng. Moreover, the complete mitogenome sequence and 10 KASP markers will be useful for ginseng research and breeding.
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Hu, Jinghuang, Jingting Li, Peipei Wu, et al. "Development of SNP, KASP, and SSR Markers by BSR-Seq Technology for Saturation of Genetic Linkage Map and Efficient Detection of Wheat Powdery Mildew Resistance Gene Pm61." International Journal of Molecular Sciences 20, no. 3 (2019): 750. http://dx.doi.org/10.3390/ijms20030750.
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The gene Pm61 that confers powdery mildew resistance has been previously identified on chromosome arm 4AL in Chinese wheat landrace Xuxusanyuehuang (XXSYH). To facilitate the use of Pm61 in breeding practices, the bulked segregant analysis-RNA-Seq (BSR-Seq) analysis, in combination with the information on the Chinese Spring reference genome sequence, was performed in the F2:3 mapping population of XXSYH × Zhongzuo 9504. Two single nucleotide polymorphism (SNP), two Kompetitive Allele Specific PCR (KASP), and six simple sequence repeat (SSR) markers, together with previously identified polymorphic markers, saturated the genetic linkage map for Pm61, especially in the proximal side of the target gene that was short of gene-linked markers. In the newly established genetic linkage map, Pm61 was located in a 0.71 cM genetic interval and can be detected in a high throughput scale by the KASP markers Xicsk8 and Xicsk13 or by the standard PCR-based markers Xicscx497 and Xicsx538. The newly saturated genetic linkage map will be useful in molecular marker assisted-selection of Pm61 in breeding for disease resistant cultivar and in its map-based cloning.
Dissertations / Theses on the topic "KBioscience's Allele-Specific PCR (KASP)"
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Benazir, Katarina Marquez. "Molecular Marker Applications in Oat (Avena Sativa L.) Breeding and Germplasm Diagnostics." Thèse, Université d'Ottawa / University of Ottawa, 2014. http://hdl.handle.net/10393/31148.
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The ability to identify germplasm and select traits accurately is fundamental to successful plant breeding. Pedigrees and molecular markers facilitate these processes; however misleading experimental results can occur when incorrect relationships and/or cultivar names are recorded. Molecular markers can identify these inconsistencies, and with advances in genotyping technology these diagnostics can be done faster and more objectively. This study aimed to develop molecular marker assays and graphical genotyping methodologies for cultivar identification, seed purity assessment and trait selection in oat (Avena sativa L.). KBioscience’s Allele-Specific PCR (KASP™) and genotyping-by-sequencing (GBS) technologies were applied to a set of current Canadian oat cultivars to evaluate their utility for identifying cultivars and detecting intra-cultivar variation. Both KASP™ and GBS detected different extents of heterogeneity among a set of 160 seeds that originated from four seed sources of four cultivars. In both cases, the detected variation did not appear to be limited to a specific cultivar or seed source, reinforcing that all cultivars are heterogeneous. Graphical genotyping localized heterogeneity to specific chromosome regions, thereby distinguishing physical contamination from true genetic heterogeneity and heterozygosity. Pre-existing genotype data for 700 oat cultivars and breeding lines were also used to construct graphical genotypes for pedigree validation and discovery of potential sources for favourable quantitative trait loci (QTL) alleles. This methodology used historical QTLs and anchoring markers to identify 25 putative “high oil” allele carriers. The results from this study will provide diagnostic tools for cultivar identification and pedigree validation, in addition to meaningful information about existing heterogeneity and possible QTL locations in current cultivars.