Back to Search
Start Over
PCR-multiplexes for a genome-wide framework of simple sequence repeat marker loci in cultivated sunflower
- Source :
- Theoretical and Applied Genetics. 107:6-19
- Publication Year :
- 2003
- Publisher :
- Springer Science and Business Media LLC, 2003.
-
Abstract
- Simple sequence repeat (SSR) and other DNA sequence-tagged site markers can be genotyped more rapidly and cost efficiently by simultaneously amplifying multiple loci (multiplex PCR). The development of PCR-multiplexes for a nearly genome-wide framework of 78 SSR marker loci in cultivated sunflower ( Helianthus annuus L.) is described herein. The most outstanding single-locus SSR markers in the public collection (300 out of 1,089) were identified and screened for polymorphisms among 24 elite inbred lines, preparatory to selecting SSR markers for testing in multiplex PCRs. The selected SSR markers produced robust PCR products, amplified a single locus each, were polymorphic among elite inbred lines (minimum, mean and maximum heterozygosities were 0.08, 0.53 and 0.85, respectively), and supply a dense genome-wide framework of predominantly or completely codominant, single-locus DNA markers for molecular breeding and genomics research in sunflower. Thirteen six-locus multiplex PCRs were developed for 78 SSR marker loci strategically positioned throughout the sunflower genome (three to five per linkage group) by identifying compatible SSR primer combinations and optimizing multiplex PCR protocols. The multiplexed SSR markers, when coupled with 17 complementary SSR marker loci, create a 'standard genotyping' set ideal for first-pass scans of the genome, as are often needed when screening bulked-segregant DNA samples or mapping phenotypic trait loci. The minimum, mean and maximum heterozygosities of the multiplexed SSR markers were 0.38, 0.62 and 0.83, respectively. The PCR-multiplexes increase genotyping throughput, reduce reagent costs, and are ideal for repetitive genotyping applications where common sets of SSR marker loci are required or advantageous.
- Subjects :
- Genetic Markers
Genetics
Polymorphism, Genetic
DNA, Plant
Chromosome Mapping
Genetic Variation
food and beverages
Genomics
General Medicine
Phenotypic trait
Biology
Polymerase Chain Reaction
Genome
Gene mapping
Genetic marker
Multiplex polymerase chain reaction
Helianthus
Microsatellite
Agronomy and Crop Science
Genotyping
Genome, Plant
DNA Primers
Microsatellite Repeats
Biotechnology
Subjects
Details
- ISSN :
- 00405752
- Volume :
- 107
- Database :
- OpenAIRE
- Journal :
- Theoretical and Applied Genetics
- Accession number :
- edsair.doi.dedup.....66122027ec2eb605775d5708334fd42a
- Full Text :
- https://doi.org/10.1007/s00122-003-1233-0