With so many variables to consider, the arduous calculations should be automated by computer software. In order to develop a good pair of PCR primers, numerous factors must be considered, including base pair length, annealing and melting temperatures, base pair repeats, etc. Well-designed primers allow the PCR system to work with accuracy and precision. PCR primers are an integral component of PCR. This has allowed for major advancements in molecular biology and precision medicine. With a minimal amount of DNA, PCR can replicate millions of copies of the targeted region. doi: 10.1007/s0012-0.Polymerase chain reaction (PCR) has been a revolutionary technology in biomedical fields. and its wild relatives: Brassica rapa L., Raphanus raphanistrum L., Sinapis arvensis L., and Erucastrum gallicum (Willd.) O.E. Hybridization between transgenic Brassica napus L. Warwick SI, Simard MJ, Legere A, Beckie HJ, Braun L, Zhu B, et al. Hybridisation within Brassica and allied genera: evaluation of potential for transgene escape. 1935 7:389–452.įitzJohn RG, Armstrong TT, Newstrom-Lloyd LE, Wilton AD, Cochrane M. napus and peculiar mode of fertilization. Genome analysis in Brassica with special reference to the experimental formation of B. Cabbage family affairs: the evolutionary history of Brassicaceae. Turnip time travels: age estimates in Brassicaceae.
The MPCR assay can be readily implemented in any basic molecular laboratory and should prove useful for the management of Brassica germplasm collections in genebanks.īrassica Brassicaceae Genebank Germplasm management Species identification Triangle of U.įranzke A, Koch MA, Mummenhoff K. In addition, the MPCR was sensitive enough to detect seed contaminations in pooled seed samples of Brassica accessions.Ī cheap and fast multiplex PCR assay for identification of Brassica species in the triangle of U was developed and validated in this study.
Further validation against 120 Brassica accessions previously genotyped showed that the MPCR is highly accurate and comparable to more advanced techniques such as SSR marker sequencing or the Illumina Infinium B. A cheaper, faster and simpler method for Brassica species identification is described here.Ī multiplex polymerase chain reaction (MPCR) consisting of new and existing primers specific to the Brassica A, B and C genomes was able to reliably distinguish all six Brassica species in the triangle of U with 16 control samples of known species identity. However, these methods are technically challenging, expensive and time-consuming, making them unsuitable for routine or rapid screening of Brassica accessions for germplasm management. Molecular genotyping based on simple sequence repeat (SSR) marker sequencing or the Illumina Infinium Brassica napus 60K single nucleotide polymorphism (SNP) array has been used to identify species and assess genetic diversity of Brassica collections. Consequently, a major issue for genebank collections is ensuring the correct identification of species. Extensive shared traits and diverse morphotypes among Brassica species make identification and classification based on phenotypic data alone challenging and unreliable, especially when dealing with large germplasm collections. The genetic relationships among the six Brassica species are described by U's triangle model. Within the Brassicaceae, six species from the genus Brassica are widely cultivated throughout the world as oilseed, condiment, fodder or vegetable crops.
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