The amplification of IGS1 region was performed in a reaction volume of 50 pL using a thermocycler.
Through a BLAST search, eight highly homologous sequences of IGS1 and seven of ITS regions were retrieved from GenBank as references material (Table 3).
The potential of integration of IGS1 and ITS regions as inter- and intraspecies marker was examined.
Secondly, by use of the same samples, the IGS1 and ITS regions gave rise to 14 and 12 unique haplotypes, respectively.
Thirdly, to recommend an efficient sequence-based DNA barcode for Pleurotus genus, the combined IGS1 and ITS regions were used.
Homology search analysis by NCBI BLAST was performed for IGS1 sequences and eight published sequences which had great similarity with the experimental strains which were detected.
In addition to the IGS1 sequences, BLAST search analysis was performed for ITS sequences by NCBI BLAST.
The obtained sequences of IGS1 and ITS regions in addition to those previously deposited into GenBank were employed to construct a combined phylogenetic tree.
The relationships between combined IGS1 and ITS haplotypes were profoundly assessed by constructing the genealogical medianjoining network (Figure 3).
gattii 10-loci MLST, for the accepted consensus MLST scheme (CAP59, GPD1, IGS1, LAC1, PLB1, SOD1, and URA5) (9), and a previously launched C.
The CAP10 locus showed the lowest overall diversity ([n.sub.ST] = 19; [D.sub.ST] = 0.765) and the IGS1 locus showed the highest diversity ([n.sub.ST] = 52; [D.sub.ST] = 0.930).