Rapid and precise genotyping of porcine microsatellites

Yue, G. H., Beeckmann, P., Bartenschlager, H., Moser, G. and Geldermann, H. (1999) Rapid and precise genotyping of porcine microsatellites. Electrophoresis, 20 17: 3358-3363. doi:10.1002/(SICI)1522-2683(19991101)20:17<3358::AID-ELPS3358>3.0.CO;2-B

Author Yue, G. H.
Beeckmann, P.
Bartenschlager, H.
Moser, G.
Geldermann, H.
Title Rapid and precise genotyping of porcine microsatellites
Journal name Electrophoresis   Check publisher's open access policy
ISSN 0173-0835
Publication date 1999-11
Sub-type Article (original research)
DOI 10.1002/(SICI)1522-2683(19991101)20:17<3358::AID-ELPS3358>3.0.CO;2-B
Volume 20
Issue 17
Start page 3358
End page 3363
Total pages 6
Place of publication Weinheim, Germany
Publisher Wiley
Language eng
Formatted abstract
Microsatellites are useful markers for genetic mapping and linkage analysis because they are highly polymorphic, abundant in genomes and relatively easily scored with polymerase chain reaction (PCR). A rapid genotyping system for microsatellites was developed, which included multiplex PCRs, multiple use of Hydrolink(TM) gels, automated fluorescent detection of fragments on an A.L.F. DNA sequencer, automatic assignment of alleles to each locus and verification of genotypes with a self-developed computer program 'Fragtest'. Eight multiplex PCRs have been developed to genotype 29 microsatellites for genetic and quantitative trait loci (QTL) mapping on pig chromosomes 6, 7, 12 and 13. Three to six microsatellites could be amplified in one multiplex PCR. Each multiplex reaction required only different concentrations of each pair of primers and a low concentration of dNTP (100 μM). A dNTP concentration of 100 μM proved to be optimal for the coamplification of microsatellites under the concentration of 1.5 mM MgCl 2. Using four internal size standards added in each sample, the 5% Hydrolink gel could subsequently be used up to five times (total running time of 500 min) on the A.L.F. automated sequencer without significant loss of resolution and precision of fragment length analysis. Automatic assignment of alleles on each locus using 'Fragtest' significantly increased the efficiency and precision of the genotyping. This system is thus a rapid, cheap, and highly discriminating genotyping system.
Keyword microsatellites
multiplex polymerase chain reaction
gel reuse
automatic genotyping
Dna Sizing Technology
Tandem Repeats
Linkage Map
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: UQ Diamantina Institute Publications
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