Candidate mutation confirmed by Sanger sequencing
The potential mutated bases and flanking sequence of genes were
amplified by polymerase chain reaction (PCR) and sequenced by Sanger
sequencing.
(R42: forward primer 5′-TACACACCAAAAATTCTGCATAGC
and reverse primer 5′-CCAGCTGAGTACGGCTACATC;
R44: forward primer 5′-CATCAGTTTGTCACTGTGTTGAGT
and reverse primer 5′-TTTGGGCTATTTGTCTTGTCATTA;
R46: forward primer 5′-GAGTCTAAGCCAGCAGGTAACAAT
and reverse primer 5′-TAGCCGGGGTATATTCTACAAGAG;
R47: forward primer 5′-TCACCCAGTTCACCACTGAG
and reverse primer 5′-ATGCAGCATAAGGCAGAAAATTG;
R51: forward primer 5′-TTGGGCTGGGTAGTAGAAAAATAG
and reverse primer 5′-GCACTGTGCTAGTGGTTTAAAAAG;
R52: forward primer 5′-GCAGGAGAGCATCTAGTTCAATC
and reverse primer 5′-CAGGAAGAAGAGCATAAGAACCTG).
All nucleotide positions were determined according to the standard gene
reference sequence.