Table 3.
Primer/probeb | Sequence (5′ to 3′)c | Size (bp) | 5′ fluorescent dye | 3′ quencher |
---|---|---|---|---|
KPC-F107 | TCGAACAGGACTTTGGCGGCT | 260 | ||
KPC-R366 | GGACAGCTCCGCCACCGTCATG | |||
MB147G | gcgct CGATGGATACCGGCTCA agcgc | FAM | DABCYL | |
MB308C | cgcga CTGGTTCCGTGGTCAC tcgcg | HEX | DABCYL | |
MB308T | cgcga CTGGTTCTGTGGTC tcgcg | Quasar 670d | BHQ-2 | |
KPC-F684 | GGCAGTCGGAGACAAAACC | 177 | ||
KPC-R860 | CCCTCGAGCGCGAGTCTA | |||
MB716T | cgcga AACCTGCGGAGTGTATGG tcgcg | Atto 425 | DABCYL | |
MB716C | cgcga AACCTGCGGAGCGTATGG tcgcg | Quasar 670d | BHQ-2 | |
MB814C | cgacg GACAAGCACAGCGAGG cgtcg | CAL Fluor Red 610 | BHQ-2 |
FAM, fluorescein; HEX, hexachlorofluorescein; DABCYL, 4-(4′-dimethylaminophenylazo)benzoic acid; BHQ, black hole quencher.
Oligonucleotide primer sets spanning nucleotide positions 107 to 366 and 684 to 860 were used to amplify two segments, each containing two molecular beacon target positions. Positions 308 and 716 are targeted by more than one molecular beacon probe.
Molecular beacon hairpin sequences are shown in lowercase letters. Single nucleotide polymorphism target positions are underlined, as in Table 2.
Quasar 670 dye is used for both MB308T and MB716C, as explained in Table 2.