Pengembangan Set Multipleks Penanda DNA Mikrosatelit untuk Analisis Variasi Genetik Padi dan Kedelai

Chaerani Chaerani, Nurul Hidayatun, Dwinita W. Utami

Abstract


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.


Keywords


Microsatellite markers; multiplex panels; fluorescently labelled M13 primer; rice; soybean.

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DOI: http://dx.doi.org/10.21082/jbio.v5n2.2009.p57-64

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P-ISSN : 1907-1094
E-ISSN : 2549-1547


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