TABLE 2.
Characteristics of primers used in this study
| Primer purpose and target gene (locus taga) | Primerb | Primer sequencec (5′-3′) | Location within genea | Annealing temp (°C) | Product size (bp) | Reference or source |
|---|---|---|---|---|---|---|
| PCR for inhA1, nprA, and hlyII detection | ||||||
| inhA1 (BC1284) | F-inhA1-d1 | ACGCITTIAAATTTGCICG | 1937-1955 | 55 | 257 | This study |
| R-inhA1-d1 | ACGCGTTGGAGATACAACTT | 2193-2174 | ||||
| F-inhA1-d2 | CCAGCTTGGAAAGTTGTATC | 2164-2183 | 55 | 166 | This study | |
| R-inhA1-d2 | CAGCTTGTCCTACTACTTCA | 2329-2310 | ||||
| nprA (BC0602) | F-nprA-d | GTATACGGAGATGGTGATGG | 1111-1130 | 55 | 263 | This study |
| R-nprA-d | GGATCACTCATAGAGCGAAG | 1373-1354 | ||||
| hlyII (BC3523) | Fhly-II | GATTCTAAAGGAACTGTAG | 94-112 | 48 | 868 | 19 |
| Rhly-II | GGTTATCAAGAGTAACTTG | 961-943 | ||||
| F-hlyII-d | CAAGTTACTCTTGATAACC | 943-961 | 48 | 194 | This study | |
| R-hlyII-dq | TCACCATTTACAAAGATACC | 1136-1117 | ||||
| PCR amplification of hlyII probe | ||||||
| hlyII (BC3523) | F-hlyII-p | GCATTTGCAGATTCTAAAGG | 85-104 | 55 | 1,019 | This study |
| R-hlyII-p | TCGTAACTGATACCATAACC | 1103-1084 | ||||
| RT-qPCR | ||||||
| inhA1 (BC1284) | F-inhA1-q | GATAAAAC(A/G)CCAGCTTGGAAAG | 2155-2176 | 60 | 189 | This study |
| R-inhA1-q | TGCAGA(A/T)TTATCATCAGCTTG | 2343-2323 | ||||
| nprA (BC0602) | F-nprA-q | TGCAGCAGCAGTAGATGCTC | 951-970 | 60 | 188 | This study |
| R-nprA-q | ATGTTACACCATCACCATC | 1138-1120 | ||||
| hlyII (BC3523) | F-hlyII-q | CTGGAAAAACCATCAAGTTACTC | 930-952 | 60 | 207 | This study |
| R-hlyII-dq | TCACCATTTACAAAGATACC | 1136-1117 | ||||
| rpoA (BC0158) | F-rpoA-q | TAACTCCTTACGTCGTATTC | 111-130 | 60 | 186 | This study |
| R-rpoA-q | ATTTCCAACGTCTTCTCTTC | 296-277 | ||||
| rpoB (BC0122) | F-rpoB-q | TCAGTGGTTTCTTGATGAGG | 111-130 | 60 | 176 | This study |
| R-rpoB-q | CTTTTACACGAAGTGGTGCT | 286-267 |
a
Reference strain Bacillus cereus ATCC 14579 (NCBI accession no. NC_004722).
b
F, forward primer; R, reverse primer.
c
I, inosine.