TABLE 1.
Primer | Sequence (5′ to 3′) | Annealing temp (°C) |
---|---|---|
FCaa | TGGGCNGGNTGYYTNGT | 53 |
FCba | TACCANGTYTGRTTCAT | 53 |
YPa | TGGAGAAAGGGAAGTTTTGCTTCTTACGAC | 67 |
YPb | GTCGTAAGAAGCAAAACTTCCCTTTCTCCA | 67 |
YNa | GGTGTGGTTTTGGGTATGAACCAGACGTGG | 67 |
YNb | GGCAATCAAGTTCATGAAGTTCTCCATGAC | 67 |
YP1 | TTCAGAAGAGGATTCAAGGGCTACAATCCG | 67 |
YP2 | CGGATTGTAGCCCTTGAATCCTCTTCTGAA | 67 |
YN1 | TGCACGGTTCTTACTATGAACCAAACCTGG | 67 |
YN2 | ACCGATAATACTCAAGAAGTTCGACATGGT | 67 |
FC1 | AGCGAAGGGTTGAGTTGC | 64 |
FC2 | ATGTCGAGTTCCGTTCTC | 64 |
FC3 | CTGGTAGAATACAACACCG | 60 |
FC4 | CGGAACGTACGAGAGAGAC | 60 |
FciSb | TCGAGAACTAGTGGCGAAACAAAGAAGAGA | 60 |
FciNc | CGAGCTATGCATACCAAACCACCAACAGAC | 60 |
Fcy2Sb | TCGAGAACTAGTGATGGAAAGGAGAGGCTG | 62 |
Fcy2Nc | CGAGCTATGCATAGATTTGCTACTCTGATG | 62 |
18S1 | TTTCGATGGTAGGATAGAGGC | 62 |
18S2 | CATGCTAACAGATTCAAGCGG | 62 |
For degenerate primers FCa and FCb, D is A, G, or T; H is A, C, or T; R is A or G; W is A or T; and Y is C or T.
The SpeI restriction sites located in primers FciS and Fcy2S are indicated in boldface type.
The NsiI restriction sites located in primers FciN and Fcy2N are indicated in boldface type.