Table 1.
Characteristics of the specific primers used in PCR
| primer | (5`- 3`) Sequence | Purpose | Annealing Temperature (°C) | Length of Production | |
|---|---|---|---|---|---|
| Primer-1 Combination | LFR1-F LFR1-R |
5' -GACAGCCTTTGGGCACTTAG-3' 5' -GGGTAGGACAGAAGCGACAG-3' |
Sequencing | 57.8 | 1068bp |
| Primer-2 Combination | HajF2 LFR1-R Lac-1 Lac-2 |
5' -TCGTTCCGGAGTCGCCCCAGGAAG-3' 5' -GGGTAGGACAGAAGCGACAG-3' 5' -GCCTCATGACAACTCCCACAC-3' 5' -CAGGTTGACACATCGGTTGAC-3' |
G allel specific detection Internal control |
65.0 - 68.0 | 408bp 301bp |
| Primer-3 Combination | HajR2 LFR1-F Lac-1 Lac-2 |
5' -GGGACGAAGAGCTTCATGGCTGTG-3' 5' -GACAGCCTTTGGGCACTTAG-3' 5' -GCCTCATGACAACTCCCACAC-3' 5' -CAGGTTGACACATCGGTTGAC-3' |
C allel specific detection Internal control |
65.0 - 68.0 | 709bp 301bp |
The underlined bases are those modified from the original sequence to increase the specificity of the allele-specific PCR.