Open AccessPengembangan Set Multipleks Penanda DNA Mikrosatelit untuk Analisis Variasi Genetik Padi dan Kedelai
Author(s) -
Chaerani Chaerani,
Nurul Hidayatun,
Dwinita Wikan Utami
Publication year - 2016
Publication title -
jurnal agrobiogen/jurnal agrobiogen
Language(s) - English
Resource type - Journals
eISSN - 2549-1547
pISSN - 1907-1094
DOI - 10.21082/jbio.v5n2.2009.p57-64
Subject(s) - multiplex , microsatellite , microbiology and biotechnology , genotype , allele , primer (cosmetics) , biology , multiplex polymerase chain reaction , genetics , chemistry , polymerase chain reaction , gene , organic chemistry
Detection of multiplex microsatellite markers in a single capillary array on a laser detection system is traditionally conducted with specific primers that are labelled with fluorescent dyes. An alternative method using fluorescent labels that are appended to 5’ end of universal primer M13 instead of to the specific primers offers flexibility in designning multiplex panels and a less expensive method. Allele size range of microsatellite loci that can be grouped in multiplex panels can be accurately estimated by pooling and analyzing DNA samples from several genotypes simultaneously. This paper describes the procedure in development of microsatellite multiplex panels using M13 fluorescentlylabelled and estimation of allele size range based on pooled DNA strategies. Two multiplex panels of PCR amplification products for rice consisting of 15 loci and three panels for soybean consisting of 10 loci have been designed. The panels have been applied to 50 accessions of rice and soybean with fairly good results. Further characterization of allele size range, however, is required prior to the application of these panels to diverse genotypes. The procedure described here should be applicable in the development of multiplex panels of other species.