TAILIEUCHUNG - Optimization of PCR amplification of wheat simple sequence repeat DNA markers

Simple Sequence Repeat (SSR) DNA markers or Microsatellites are di-, tri-, tetra-nucleotide tandem repeats containing loci of eukaryotic genomes. It is demonstrated that these loci are very polymorphic due to the change in the number of repeating units among the individuals of populations. | Turk J Biol 25 (2001) 153-158 © TÜB‹TAK Optimization of PCR Amplification of Wheat Simple Sequence Repeat DNA Markers Nurdan DO⁄RAR, Mahinur S. AKKAYA Middle East Technical University, Department of Chemistry, Biochemistry Division, TR-06531, Ankara - TURKEY Received: Abstract: Simple Sequence Repeat (SSR) DNA markers or Microsatellites are di-, tri-, tetra-nucleotide tandem repeats containing loci of eukaryotic genomes. It is demonstrated that these loci are very polymorphic due to the change in the number of repeating units among the individuals of populations. Each SSR locus can easily be amplified by using a polymerase chain reaction (PCR) knowing the DNA sequence flanking the repeat region specifically. The only limiting feature of the application of these markers is the need for prior sequence information for developing primers in locus-specific PCR amplification. This limitation is alleviated for the economically important species and the ones closely related, since primer sequences of the SSR DNA markers and the amplification conditions are available in the literature. However, when the reported PCR amplification conditions were applied, with the 2 wheat SSR markers, we found that contaminating bands appeared on the gels. This is most likely due to different laboratory conditions. In this study, we report the optimization of PCR conditions for WMS30 and WMS46 of wheat. The most important parameters were found to be the annealing temperature and Mg2+ ion concentrations. Key Words: Triticum durum, microsatellites, Simple Sequence Repeat DNA (SSR) markers, differentiation, DNA fingerprinting Bu¤day Basit Dizilim Tekrar DNA Belirleyicilerinin PZR Ço¤altma Optimizasyonu Özet: Basit Dizilim Tekrar (BDT) DNA belirleyicileri ya da Mikrosatellitler, ökaryot genomlar›nda bulunan di-, tri-, tetra-nükleotit tekrar üniteleridir. Bu belirleyici lokuslar›n›n popülasyonlar›n bireyleri aras›nda oldukça polimorfik olmalar›, tekrar ünitelerinin say›lar›ndak› .

• Sinh học
• Triticum durum
• Simple sequence repeat DNA markers
• DNA fingerprinting
• DNA sequence flanking
• Eukaryotic genomes
• Durum wheat
• Genetic advance
• Wheat (Triticum durum Desf
• Genetic advance in durum wheat
• Genetic variability
• Amino acids
• Irrigation conditions
• Durum wheat (Triticum durum L
• Variation in lysine content
• Genotype × environment interaction
• Grain vitreousness percentage
• Wheat Triticum durum L
• Grain yields
• Spot blotch
• Path analysis
• Disease severity
• Area under disease progress curve
• Durum wheat (Triticum durum L )
• In situ chromosome
• Aegilops tauschii
• Triticum Durum X
• In situ chromosome doubling method
• Imperata cylindrica L
• Interspecific cross
• Triticum durum L
• High quality durum wheat
• Wheat cultivars Triticum durum L
• Ascorbic acid
• Durum wheat cultivars
• Non protein SH groups
• Durum wheat Triticum durum genotypes
• Boron toxicity
• Boron concentration
• Genotypic variation
• Sowing dates
• Yield and sub tropical zone
• Sub tropical zone of Jammu
• Performance of durum wheat
• Aegilops sp
• Polymorphic AFLP markers
• Polymerase chain reaction
• BMC Genomics
• Triticum turgidum durum
• De novo assembly
• Triticum aestivum L
• Stripe rust
• Leaf rust
• Triticum aestivum
• Tetraploid wheat
• STR marker
• Tetraploid durum wheat
• γ gliadin 45
• LMW 2 glutenin
• Backcross breeding
• Marker assisted selection
• Wheat landraces
• Polyphenol oxidase
• Sodium dodecyl sulfate
• Acidic polyacrylamide gel electrophoresis
• Determining some yield
• Mutation breeding
• Yellowberry kernel
• Agronomic potential
• Triticum turgidum var
• durum Desf
• Grain yield
• Genotype–environment
• Agro ecological
• Flag leaf area
• Morphological parameters
• Plant height
• Specific leaf weight
• Path coefficient analysis
• Correlation for yield
• Components traits in durum wheat
• Grain yield per plot
• BMC Genetics
• Local landraces
• Landrace characterization
• Phenotypic diversity
• Genetic diversity
• Population structure
• Ethiopian durum wheat
• Improved varieties
• Wheat Triticum durum
• Chromosomal abnormalities
• Mitotic abnormalities
• Chromosomal behavior
• Wheat seeds
• Phenotypic stability analysis
• Macaroni wheat
• Macaroni wheat (Triticum durum desf )
• Stress environments
• Experimental material comprised
• Durum and soil fertility
• Response of wheat varieties
• Wheat varieties
• Temperature situation
• Various phenophases of wheat crop
• Bread Triticum aestivum L
• Boron deficiency
• Boron fertilization
• Wheat cultivars
• Linkage disequilibrium
• Rare alleles
• Triticum turgidum L