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Specific S. divinorum primers were designed on the sequence of the 5S-rRNA gene spacer region.
PCR primers were designed on the sequence of TaLCYB using the Primer 5.
For CAPS (cleaved amplified polymorphic sequence) marker development, PCR primers were designed on the sequence of the A17 genotype.
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Primers designed on the sequences of the first DNA region for the virB4 (Oant_0677), virB5 (Oant_0678) and virB8 (Oant_0682) orthologs gave a positive amplification from O. anthropi genomic DNA and cDNA indicating their expression (Fig. 1D).
Species-specific primers were designed on the sequences of the first successfully amplified and sequenced fragments, and used to amplify the remaining regions of the genome.
To confirm that each of the assembled cDNA sequences originated from a single, full-length cDNA, primers were designed on the sequences of the 5' and 3' untranslated regions (UTRs).
If necessary, species-specific primers were designed on the sequences of the initially obtained amplicons, and used to amplify the remaining portion(s) of the mt genome.
As shown in Figure 1B, we used different forward primers (F1 to F8) designed on the genomic sequence and coupled them to REV 3 in PCR reactions.
A reverse primer (R) was then designed on the obtained sequence.
Primers were designed on the consensus sequence of the contigs, in order to amplify the region containing the SNP.
Its related cDNA was obtained by RT-PCR experiments using specific oligonucleotides designed on the X60760 sequence [ 3].
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