Table 2.
Table 2A. Primer sequences used for PCR amplification and sequencing | ||
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| ||
Primer | Primer Sequence 5′-3′ | PCR Product Size |
12Sai | AAACTAGGATTAGATACCCTATTA T | ∼400 base pairs |
12Sbi | AAGAGCGACGGGCGATGTGT | |
16Sa | CGCCTGTTTATCAAAAACAT | ∼400 base pairs |
16Sb | CTCCGGTTTGAACTCAGATCA | |
COIF | TAAACTTCAGGGTGACCAAAAAATCA | ∼ 600 base pairs |
COIR | GGTCAACAAATCATAAAGATATTGG | |
28Sa | GACCCGTCTTGAAACACGGA | ∼ 600 base pairs |
28Sb | TCGGAAACCAGCTAC |
Table 2B. PCR amplification profiles | ||||||
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Loci | Number of Cycles | Hotstart | Denaturation | Annealing | Extension | Final Extension |
12S | 25 | 94°C for 2 min | 94°C for 15 sec | 42°C for 30 sec | 72°C for 30 sec | 72°C for 7 min |
16S | 25 | 94°C for 2 min | 94°C for 15 sec | 42°C for 30 sec | 72°C for 30 sec | 72°C for 7 min |
CO1 | 25 | 94°C for 2 min | 94°C for 15 sec | 36°C for 30 sec | 72°C for 30 sec | 72°C for 7 min |
28S | 25–30 | 94°C for 2 min | 94°C for 15 sec | 40–49°C for 30 sec | 72°C for 30 sec | 72°C for 7 min |