Table 1 Used primers, amplification programme and reaction mixtures
Oligonucleotide Sequence (5’- > 3’) | Reaction mixture | Amplification Program | ||
|---|---|---|---|---|
F: ACGGCCNNRACTCCTAC R: TTACGGNNTGGACTACHV | Water | 2.6 is µl | 95 ºC | 5 min |
Kappa | 5.0 µl | 95 ºC | 15 s (× 40) | |
Primer F (10 µM) | 0.2 µl | 48 ºC | 20 s (40x) | |
Primer R (10 µM) | 0.2 µl | 72 ºC | 30 s (40 s) | |
DNA | 2.0 µl | 72 \(^\circ C\) | 5 min | |
F: CCTACGGGNGGCWGCAG R: GACTACHVGGGTATCTAATCC | Water | 10.5 µl | 95 ºC | 5 min |
Kappa | 12.5 µl | 95 ºC | 30 s (25x) | |
Primer F (10 µM) | 0.5 µl | 55 ºC | 30 s (25x) | |
Primer R (10 µM) | 0.5 µl | 72 ºC | 30 s (25x) | |
DNA | 1.0 µl | 72 ºC | 5 min | |