TABLE 2.

PCR primers and conditions used to amplify the designated M. tuberculosis gene regionsa

Gene regionForward primerReverse primerSize (bp)Length of D(s)A (temp [°C], time [s])Length of E(s)
furA 5′-GCGATCGGGTCCTAGCAG-3′5′-TTCATATGACCCACGACGG-3′6414061.0, 3040
Rv0340 5′-ATGCGTCGTATGCTTGG-3′5′-CCAAACACCTATCGGGATC-3′8504058.5, 3040
iniB 5′-ATAAGTTCCGGACCGGCG-3′5′-CGACAGATGAGGCATAGCAG-3′1,0534056.0, 3045
iniB-iniA 5′-TTGAACGGCGCTGCTATG-3′5′-GTGCTGATGTCATCGACGG-3′1,0704062.0, 3045
iniA 5′-CAACCGCAGCGGTTGACAT-3′5′-CCGCATGCCGATAATCATT-3′1,0794061.0, 3045
iniA-iniC 5′-GGAATCGAAACCGCTGCG-3′5′-CCAGCCCACCGATCTGTTTGA-3′1,0904064.0, 3045
iniC 5′-TCCTGTTGCGCACCCTGAAC-3′5′-AACATGTTCCACCCGGTGGC-3′1,0404065.0, 3045
srmR homolog5′-GCCAGTACCGGATCGACG-3′5′-GTGTCGCGGACATCCTGG-3′8193063.5, 3040
5′-CCAGGATGTCCGCGACACC-3′5′-TGTCCGGGTGCGAGCAAC-3′8353067.5, 3040
fabD 5′-AAAAACATAGCTTACAGGCCCG-3′5′-GTTGTGTACAAATCGAACTGACG-3′1,0714059.5, 3045
kasA 5′-GTTCAGGCGAGGCTTGAG-3′5′-GCGATGTCGTGCTTCAGTAA-3′1,2934560.5, 3050
accD6 5′-GACAGGAGACCTGCGATGAC-3′5′-GGCAGAACAATCCGACCA-3′6553061.5, 3030
5′-CGGTTGGTCGGATTGTTC-3′5′-CCTCGCGCGTGCGATTCTG-3′8423561.0, 3035
fadE24 5′-GCTATCAGATGCCTGGCG-3′5′-GTGGGATCGAATAGTGGCTG-3′8903561.0, 3035
fbpC 5′-GCATGGGTCTCTCCTCTG-3′5′-GCTGATACCAGTCGGTGTAG-3′6353056.0, 3030
5′-GCCAATCCAGTTTCTACACCG-3′5′-ATGTTCCACACATCGGC-3′7323557.5, 3035
efpA 5′-ACCTCCCGCGATCATCG-3′5′-CGTCGAGCTTCATCCGTTC-3′8103564.5, 3035
Rv1592c 5′-AACTCGGCGTACCCAACC-3′5′-ACACGCTCGGAATTCAAGG-3′8043561.5, 3035
5′-GAGCGTGTCGGGTTGTCC-3′5′-CGAGGTTGTGTGCCAGGTC-3′8343564.0, 3035
Rv1772 5′-CGGGTGTTTCTCAACGAC-3′5′-GGACTGGACTCGCTGATTG-3′6333057.5, 3030
ndh 5′-ATCACCACCGCCGCTGAAGC-3′5′-GTTCGGGTACCCGGGAATG-3′1,1344565.0, 3050
5′-CATTCCCGGGTACCCGAAC-3′5′-GTCGACCGTTTTGGCGTTGG-3′5353065.0, 3030
nhoA 5′-CTAGTGGCGCGAGCAGAC-3′5′-CCGACGTCGACGAGATAGC-3′9274061.5, 3040
5′-GCTATCTCGTCGACGTCG-3′5′-GCATTCTACGTCTACGCCG-3′6623059.0, 3030
  • a Abbreviations: D, denaturation at 94°C; A, primer annealing conditions; and E, extension at 72°C. All PCR were run for 25 cycles, were preceded by a denaturation step at 95°C for 15 min, and included a final extension step at 72°C for 7 min.