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Sequencing and identification of mutations
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The amplified fragments were sequenced in two directions (forward and reverse), using a genetic analyzer (ABI 3130 Applied Biosystem®) and the Big Dye Terminator methodology (Applied Biosystem®), following standardized conditions at Vanderbilt Genetic Institute Core Facilities, United States. The edition and alignment of the sequences was carried out using Bioedit software V 7.1.11® (Hall, 1999). Changes in sequences were matched by local alignment, with the reference sequence for 23S rRNA gene, code GenBank: U27270.1[8].
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