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Further, marker sequences are not randomly dispersed, especially for sequence repeats such as SSRs.
In meiotic linkage mapping, relationships among polymorphic marker sequences are determined by relative frequency of recombination.
Marker sequences are available in Table S1.
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Genome supercontigs containing the RFLP marker sequences were downloaded from VectorBase and screened with Tandem Repeats Finder (TRF) software using the default parameters [25].
Each of the 148 DArT marker sequences were queried against 1) all other sequenced DArT markers in the same study and 2) GenBank (www.ncbi.nlm.nih.gov) to identify putative sequence identities using the Basic Local Alignment Search Tool [23] for nucleotides (blastn) and translated to proteins (tblastx).
Orthologs for a majority of Ambystoma marker sequences were identified in more than one reference genome.
Inversions of marker sequences were filtered out by discarding inconsistent loci with the exception of very closely linked markers.
Here, C3 marker sequences were defined as the sequences that all C3 species were consistent (without considering F. pringlei), and C3-C4 marker sequences were defined as those sequences that all C3-C4 species were consistent (without considering F. angustifolia).
The ordered marker sequences were confirmed by the 'Ripple' command and finally the linkage groups were generated by 'Map' command.
In both cases, the (proxy) marker sequences were aligned to the DM assembly using BLAST, adopting stringent matching criteria.
The marker sequences were applied to the BLAST search of the reference genome sequence [ 38] to identify the specific scaffold for each marker (Table 4).
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