TABLE 5.
Primers used for TMB-1 site-directed mutagenesis and sequencinga
| Primer name | Forward primer sequence (5′–3′) | Tm (°C) | Reverse primer sequence (5′–3′) | Tm (°C) |
|---|---|---|---|---|
| TMB-1 E119Q | ACCCATAGCCATCAAGATAAAACCGCA | 74.7 | TGCGGTTTTATCTTGATGGCTATGGGT | 74.7 |
| TMB-1 E119S | ACCCATAGCCATTCAGATAAAACCGCA | 74.7 | TGCGGTTTTATCTGAATGGCTATGGGT | 74.7 |
| TMB-1 E119A | ACCCATAGCCATGCAGATAAAACCGCA | 76.2 | TGCGGTTTTATCTGCATGGCTATGGGT | 76.2 |
| M13 | GTAAAACGACGGCCAGT | 47.1 | GGAAACAGCTATGACCATG | 48.9 |
a
Mutated nucleotides are underlined. Melting temperatures were calculated at http://www.biophp.org/minitools/melting_temperature/demo.php.