caa a22 4500 475794524 CHVBK 20180406123720.0 cr unu---uuuuu 170329e20000701xx s 000 0 eng 10.1007/s001220051458 doi (NATIONALLICENCE)springer-10.1007/s001220051458 Use of 5´-anchored primers for the enhanced recovery of specific microsatellite markers in Brassica napus L [Elektronische Daten] [J. P. Varghese, B. Rudolph, M. I. Uzunova, W. Ecke] Abstract : Microsatellite markers have assumed great significance in biological research. The isolation and characterisation of microsatellites involves DNA library construction and screening, DNA sequencing, primer design and PCR optimisation. When a microsatellite is situated close to the beginning or end of a cloned fragment, specific primers cannot be designed for one of the flanking sequences, thus hindering the utilisation of such microsatellites as markers. The present approach was to use one 5´-anchored primer complementary to the microsatellite sequence in combination with one specific Cy5- labelled primer with a view to retrieving useful microsatellites, which would otherwise be lost. Six pairs of a 5´ anchored primer and a specific primer were used across a set of 31 Brassica napus winter cultivars and one accession each of five additional Brassica species. Using laser fluorometry a single labelled product was observed after amplification with each of four primer pairs, and one primer pair gave two labelled products. Three products corresponded in size with the products expected if 5´ anchoring was effective, indicating the amplification of locus-specific full-length products including all of the microsatellite repeats. All six primer pairs showed polymorphisms across the Brassica species examined, but only one primer pair showed polymorphisms within B. napus, making it useful for genetic analysis in rapeseed cultivars. The other primer pairs could be useful in studying gene introgression into B. napus or for investigating interspecific crosses involving different Brassica species. Springer-Verlag Berlin Heidelberg, 2000 Key words Anchored primers nationallicence Microsatellites nationallicence Simple sequence repeats nationallicence Polymorphism nationallicence PCR nationallicence Varghese J. P. Department of Botany, CMS College, Kottayam 686001, Kerala, India, IN aut Rudolph B. Institut für Pflanzenbau und Pflanzenzüchtung, Georg-August-Universität, Von-Siebold-Straße 8, D-37075 Göttingen, Germany, Present address: M.I. Uzunova, Kleinwanzlebener Saatzucht AG, PO 1463, D-37555 Einbeck, Germany, DE aut Uzunova M. I. Institut für Pflanzenbau und Pflanzenzüchtung, Georg-August-Universität, Von-Siebold-Straße 8, D-37075 Göttingen, Germany, Present address: M.I. Uzunova, Kleinwanzlebener Saatzucht AG, PO 1463, D-37555 Einbeck, Germany, DE aut Ecke W. Institut für Pflanzenbau und Pflanzenzüchtung, Georg-August-Universität, Von-Siebold-Straße 8, D-37075 Göttingen, Germany, Present address: M.I. Uzunova, Kleinwanzlebener Saatzucht AG, PO 1463, D-37555 Einbeck, Germany, DE aut https://doi.org/10.1007/s001220051458 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s001220051458 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Varghese J. P. Department of Botany, CMS College, Kottayam 686001, Kerala, India, IN aut NATIONALLICENCE 700 1- Rudolph B. Institut für Pflanzenbau und Pflanzenzüchtung, Georg-August-Universität, Von-Siebold-Straße 8, D-37075 Göttingen, Germany, Present address: M.I. Uzunova, Kleinwanzlebener Saatzucht AG, PO 1463, D-37555 Einbeck, Germany, DE aut NATIONALLICENCE 700 1- Uzunova M. I. Institut für Pflanzenbau und Pflanzenzüchtung, Georg-August-Universität, Von-Siebold-Straße 8, D-37075 Göttingen, Germany, Present address: M.I. Uzunova, Kleinwanzlebener Saatzucht AG, PO 1463, D-37555 Einbeck, Germany, DE aut NATIONALLICENCE 700 1- Ecke W. Institut für Pflanzenbau und Pflanzenzüchtung, Georg-August-Universität, Von-Siebold-Straße 8, D-37075 Göttingen, Germany, Present address: M.I. Uzunova, Kleinwanzlebener Saatzucht AG, PO 1463, D-37555 Einbeck, Germany, DE aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer