Leroy et al. (2000) used 4 microsatellite primers to characterise Brassica oleracea accessions. Among the 136 reproducible fragments generated, 25 (18.4%) fragments were common for all Brassica, 27 (19.9%) were unique and 84 (61.7%) were phylogenetically informative. Flannery et al. (2006) assessed polymorphisms in Brassica, Arabidopsis, Camelina, Raphanus and Sinapis using 10 plastid SSR primer sets. Eight loci were polymorphic, and separated the individuals of Brassicaceae into taxon-specific groups (Arabidopsis, Camelina, Sinapis and Brassica genera). Louarn et al. (2007) also evaluated 59 Brassica oleracea cultivars for microsatellite polymorphisms. All SSR markers, except one, produced a polymorphic information content (PIC value) of 0.5
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The same reaction mixture without genomic DNA was set up to serve as a negative control. The PCR reactions were then amplified in a DNA thermocycler (G-Storm GS1 thermal cycler, GRI Ltd., Gene House, Essex, United Kingdom) programmed as follows: (1) 94°C for 2 min (initial denaturation); (2) 94°C for 1 min (denaturation), (3) 56°C (in the case of Ol12-E03 primer) or 52°C (in the case of for Ol12-F11, Ol10-H02, Ol10-A03a, Ol10-F11a primers) for 1 min (annealing), 72°C for 1 min (extension) per 35 cycles; (4) 72°C for 20 min (final extension) then 4°C (infinitive). The amplified PCR products were resolved on 2% (w/v) agarose gels stained with ethidium bromide. A 50 bp DNA ladder was used as a DNA molecular weight standard. Bands were detected and photographed using a UVP gel documentation system (Ultra-Violet Products Ltd., Cambridge, United …show more content…
Moreover, the effective number of alleles per locus (Ae) varied from 1.597 in the accession of cabbage HRIGRU4497 to 2.394 in the accession of spring cabbage HRIGRU4566, with a mean of 1.906 (Table 3). The mean number of alleles per locus (A) ranged from 1.8 to 2.8 with an average of 2.096. These results indicated that the SSR loci in Brassica oleracea accessions analysed presented uneven allele frequencies (Ae=
The same reaction mixture without genomic DNA was set up to serve as a negative control. The PCR reactions were then amplified in a DNA thermocycler (G-Storm GS1 thermal cycler, GRI Ltd., Gene House, Essex, United Kingdom) programmed as follows: (1) 94°C for 2 min (initial denaturation); (2) 94°C for 1 min (denaturation), (3) 56°C (in the case of Ol12-E03 primer) or 52°C (in the case of for Ol12-F11, Ol10-H02, Ol10-A03a, Ol10-F11a primers) for 1 min (annealing), 72°C for 1 min (extension) per 35 cycles; (4) 72°C for 20 min (final extension) then 4°C (infinitive). The amplified PCR products were resolved on 2% (w/v) agarose gels stained with ethidium bromide. A 50 bp DNA ladder was used as a DNA molecular weight standard. Bands were detected and photographed using a UVP gel documentation system (Ultra-Violet Products Ltd., Cambridge, United …show more content…
Moreover, the effective number of alleles per locus (Ae) varied from 1.597 in the accession of cabbage HRIGRU4497 to 2.394 in the accession of spring cabbage HRIGRU4566, with a mean of 1.906 (Table 3). The mean number of alleles per locus (A) ranged from 1.8 to 2.8 with an average of 2.096. These results indicated that the SSR loci in Brassica oleracea accessions analysed presented uneven allele frequencies (Ae=