Region | PCR primers | Amplification conditions | Amplicon sizea |
---|---|---|---|
Fragment 1 | |||
First Reaction | F:5′-GCA TCT AAA TGG CTA CTG TGA TAT AC-3′ | 94 °C 5 min (1×); 94 °C 1 min, 53 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 595 bp |
R:5′-GGC ATC TGT AAA TAG ACC ATC C-3′ | |||
Nested Reaction | F:5′-TAA TGA AAA GGA ATT AGA AAT G-3′ | 94 °C 5 min (1×); 94 °C 1 min, 43 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 533 bp |
R:5′-CCA TAT TGA ATA CTA TAT TTT TC-3′ | |||
Fragment 2 | |||
First Reaction | F: 5′-CCA TAC TAT TTA ATT GCA CTA AAG-3′ | 94 °C 5 min (1×); 94 °C 1 min, 58 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 723 bp |
R:5′-ACA CAC ATA AGC AGT TTT CAA AAA G-3′ | |||
Nested Reaction | F:5′-AGG GAA ATT GAT AAA TTA AAT ATT TC-3′ | 94 °C 5 min (1×); 94 °C 1 min, 58 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 666 bp |
R:5′-CTT TTC AAG ATA TTG CAA TTT GGA-3′ | |||
Fragment 3 | |||
First Reaction | F:5′-GCA ATG TTC TAC AAA ATC AGT ACA AAG-3′ | 94 °C 5 min (1×); 94 °C 1 min, 53 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 765 bp |
R:5′-CTA CAA AAG CTG CTA GTA CAT GTC T-3′ | |||
Nested Reaction | F:5′-GAA GAA GAA TGA CAA ACT TAA AAA C-3′ | 94 °C 5 min (1×); 94 °C 1 min, 43 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 632 bp |
R:5′-CTT TTA AAA TAC TAT ATT CTT TAA TAT G-3′ | |||
Fragment 4 | |||
First Reaction | F:5′-GAG GTG ATC GTG TTT CCC ATT G-3′ | 94 °C 5 min (1×); 94 °C 1 min, 48 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 532 bp |
R:5′-CAT TTT CAG TTT TTT TAT CAC CC-3′ | |||
Nested Reaction | F:5′-GTA AGA AAG AAA AGG AAA ATC CAT TAG-3′ | 94 °C 5 min (1×); 94 °C 1 min, 49 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 443 bp |
R:5′-CTG AGT CTT GTA CAA CTT GGT C-3′ | |||
Fragment 5 | |||
First Reaction | F:5′-GAC CAA GTT GTA CAA GAC TCA G-3′ | 94 °C 5 min (1×); 94 °C 1 min, 48 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 737 bp |
R:5′-GTA AGT TAA ACA TAA TTA ATA AAG CTG-3′ | |||
Nested Reaction | F: 5′-GGG TGA TAA AAA AAC TGA AAA TG-3′ | 94 °C 5 min (1×); 94 °C 1 min, 49 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 631 bp |
R:5′-GCC GAG GAA ACT TGA AGA AC-3′ |