caa a22 4500 397547447 CHVBK 20180308164742.0 cr unu---uuuuu 161202e19960101xx s 000 0 eng 10.1093/aesa/89.1.20 doi (NATIONALLICENCE)oxford-10.1093/aesa/89.1.20 Identification of Mitochondrial DNA of Apis mellifera (Hymenoptera: Apidae) Subspecies Groups by Multiplex Allele-Specific Amplification with Competing Fluorescent-Labeled Primers [Elektronische Daten] [Myeong-Lyeol Lee, H. Glenn Hall] Allele-specific amplification (ASA) tests are described that facilitate mitochondrial DNA (mtDNA) identification of 3 major groups of honey bee, Apis mellifera L., subspecies: African, west European, and east European. At each of 3 polymorphic regions, 1 of 2 allelic sequences (representing the presence or absence of a restriction enzyme site) is specific to a different subspecies group. For each region, 3 primers are synthesized: 2 that differ by 1 nucleotide at the 3′ end, representing the 2 allelic forms, and 1 that serves as a common primer for the distal end of the region to be amplified. In 1 type of ASA test, the allelic primers are used in separate polymerase chain reactions, together with the common primer, for each of the 3 regions. Significant amplification is achieved only in those reactions that have primers that match the mtDNA sequence, thereby identifying the mtDNA type. In another type of ASA test, the allelic primers are differentially labeled with fluorescent dyes, and both are included as competitors in the same reaction along with the common primer. The primers that match the mtDNA sequences preferentially support amplification, and the colors of the amplified products identify the mtDNA type. The 3 polymorphic regions of honey bee mtDNA can be amplified in a single multiplex reaction. Advantages and limitations of ASA tests, as well as instrumentation and modifications which would enable automation, are discussed. © 1996 Entomological Society of America Systematics nationallicence honey bee subspecies nationallicence identification nationallicence mitochondrial DNA nationallicence polymerase chain reaction nationallicence allele-specific amplification nationallicence fluorescent-labeled primers nationallicence Lee Myeong-Lyeol Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611. aut Hall H. Glenn Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611. aut Annals of the Entomological Society of America Oxford University Press 89/1(1996-01-01), 20-27 0013-8746 89:1<20 1996 89 aesa https://doi.org/10.1093/aesa/89.1.20 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/aesa/89.1.20 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lee Myeong-Lyeol Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611 aut NATIONALLICENCE 700 1- Hall H. Glenn Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611 aut NATIONALLICENCE 773 0- Annals of the Entomological Society of America Oxford University Press 89/1(1996-01-01), 20-27 0013-8746 89:1<20 1996 89 aesa Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford