PCR Reaction | RFLP Reaction | ||||||
---|---|---|---|---|---|---|---|
Reaction | Primer Sequence (5′→3′) | Ann (Time) | Size | Position | Restriction Enzyme | Product Size | |
(bp) | Â | (bp) | |||||
Nest-1A | F: TTGGTAATAAAACTCCCGC | 58 (2’) | 948 | - | - | - | - |
R: TATTCCTCTTAGCACCACTCC | |||||||
Nest-2A | F: TCATCTACCGCTATTGTATG | 60 (1’) | 775 | L263E | Apo I | L: 775 | E: 633+142 |
R: TCCTCTTAGCACCACTCC | E431K | MBoII | E: 299+241+117+69+49 | K: 416+241+69+49 | |||
Nest-1B | F: AAGAAGGATAAATCACCAAG | 55 (1’) | 725 | - | - | - | - |
R: AAATACACGTATACCAGCC | |||||||
Nest-2Ba | F: TAACCATTCTAATTATACTACAGCg CAGG | 60 (1’) | 141 | A623E | Cac8 I | A: 114+27 | E: 141 |
R: TGTGTTGATGTGGTATTTATTTTATTACCC | Â | Â | Â | Â | Â | Â | |
Nest-2Bb | F: AGAACAtTTAGCTTTGCTTATAAAAAAc TAA | 60 (1’) | 164 | S769N | DdeI | S: 136+28 | N: 164 |
R: ATATGGCATAATCTAATTGCTCTTCCTAC |