HA Primer pairs and PCR conditions for DHPLC analysis | |||||||
---|---|---|---|---|---|---|---|
Exon primer | Sequence (5' to 3') | Length of PCR amplicon (bp) | Annealing Temp. (°C) | DHPLC oven (°C) | Elution profile(B%) | ||
1F | ACATCCAGTGGGTAAAGTTC | 356 | 53 | 60 | 55 | 56–65 | 52–61 |
1R | AGACTTACATCCCCACAATC | Â | Â | Â | Â | Â | Â |
2F | TTGAAGTGTCCACCAAAATGAACGACT | 211 | 54 | 59 |  | 48–57 |  |
2R | GATACCCAATTTCATAAATAGCATTCA | Â | Â | Â | Â | Â | Â |
3F | GTACTATCCCCAAGTAACCTT | 204 | 54 | 59 |  | 50–59 |  |
3R | CATAGAATGACAGGACAATAGG | Â | Â | Â | Â | Â | Â |
4F | TACAGTGGATATAGAAAGGAC | 296 | 54 | 57 |  | 54–63 |  |
4R | TGCTTATTTCATCTCAATCCTACGCTT | Â | Â | Â | Â | Â | Â |
5F | CCTCCTAGTGACAATTTCCTA | 188 | 54 | 55 |  | 49–58 |  |
5R | AGCAGAGGATTTCTTTCAGGAATCCAA | Â | Â | Â | Â | Â | Â |
6F | CATGAGACACCATGCTTAGCT | 224 | 54 | 60 |  | 51–60 |  |
6R | AACTCTGGTGCTGAATTTGGAAGACCCT | Â | Â | Â | Â | Â | Â |
7F | CAGATTCTCTACTTCATAGCCATAG | 324 | 54 | 58 | 56 | 55–64 | 53–62 |
7R | ATTAAAAGTAGGACTGGATA | Â | Â | Â | Â | Â | Â |
8F | ATATAGCAAGACACTCTGACA | 338 | 54 | 58 |  | 55–64 |  |
8R | AGAGAGTACCAATAGTCAAA | Â | Â | Â | Â | Â | Â |
9F | AGAGTTGGATTTGAGCCTACC | 284 | 54 | 58 | 55 | 53–62 | 50–59 |
9R | CAGACTTTTTCTTCTTACCTGACCTT | Â | Â | Â | Â | Â | Â |
10F | GGATTTGATCTTAGATCTCGC | 204 | 53 | 56 | 54 | 50–59 | 48–57 |
10R | ATTTTAGTTGTTATTGATGA | Â | Â | Â | Â | Â | Â |
11F | TTGAGCTATTTATGGTTTTG | 294 | 53 | 58.5 | 56 | 53–62 | 51–60 |
11R | GACATACACTGAGAATGAA | Â | Â | Â | Â | Â | Â |
12F | GCATTTCTTTACCCCTTTCA | 230 | 54 | 59.5 | 57 | 51–60 | 49–58 |
12R | CTTTATTCACCACCCACTG | Â | Â | Â | Â | Â | Â |
13F | TCCTGGGAATAAGATAATGG | 393 | 54 | 57.5 | 56 | 56–65 | 54–63 |
13R | AGAGCATACGAATGGCTAGT | Â | Â | Â | Â | Â | Â |
14(I)F | ATCTGTGTTATGAGTAACCA | 430 | 54 | 57.5 |  | 57–66 |  |
14(I)R | TCATATTTGGCTTCTTGGAG | Â | Â | Â | Â | Â | Â |
14(II)F | CATGGGCTATCCTTATCTGA | 479 | 54 | 56.5 |  | 57–66 |  |
14(II)R | CATGAACTTTCTTGGCTATT | Â | Â | Â | Â | Â | Â |
14(III)F | TCAAAGTTGTTAGAATCAGG | 441 | 54 | 55.5 |  | 56–65 |  |
14(III)R | ATTTTGTGCATCTGGTGGAA | Â | Â | Â | Â | Â | Â |
14(IV)F | GTCCAACAGAAAAAAGAGGG | 481 | 54 | 57 | 54.5 | 58–67 | 55–64 |
14(IV)R | CTACATTTTGCCTAGTGCTC | Â | Â | Â | Â | Â | Â |
14(V)F | CTGGCACTAAGAATTTCATG | 429 | 54 | 57.5 | 56 | 57–66 | 55–64 |
14(V)R | CCTTCTCATTGTAGTCTATC | Â | Â | Â | Â | Â | Â |
14(VI)F | GAAACATTTGACCCCGAGCA | 431 | 54 | 57.5 |  | 56–65 |  |
14(VI)R | TTTTGGGCAAGTCTGGTTTC | Â | Â | Â | Â | Â | Â |
14(VII)F | CACATACAAGAAAGTTGAGA | 436 | 54 | 59 | 57.5 | 56–65 | 55–64 |
14(VII)R | CTCATTTATTGCTGCTATTG | Â | Â | Â | Â | Â | Â |
14(VIII)F | GATACCATTTTGTCCCTGAA | 415 | 54 | 57.5 |  | 56–65 |  |
14(VIII)R | GTCACAAGAGCAGAGCAAAG | Â | Â | Â | Â | Â | Â |
15F | CACCTAGGAAAATGAGGATGT | 300 | 53 | 58.5 | 56 | 53–62 | 51–60 |
15R | ATAGTCAGCAAGAAAATAAA | Â | Â | Â | Â | Â | Â |
16F | AAGATCCTAGAAGATTATTC | 330 | 50 | 57.5 |  | 53–62 |  |
16R | TTAGTACACAAAGACCATTT | Â | Â | Â | Â | Â | Â |
17F | TGATGAGAAATCCACTCTGG | 349 | 54 | 58 | 56.5 | 54–63 | 53–62 |
17R | GTGCAATCTGCATTTCACAG | Â | Â | Â | Â | Â | Â |
18F | GTGGAATCCTCATAGATGTCA | 312 | 53 | 57 |  | 54–63 |  |
18R | GAGTAGGTAGAAGAAAGAGCAC | Â | Â | Â | Â | Â | Â |
19F | GCAAGCACTTTGCATTTGAG | 305 | 52 | 59 | 56 | 55–64 | 52–61 |
19R | AGCAACCATTCCAGAAAGGA | Â | Â | Â | Â | Â | Â |
20F | CCATTTTCATTGACTTACATTTGAG | 193 | 53 | 59.5 | 56.5 | 49–58 | 46–55 |
20R | AGATATAATCAGCCCAGGTTC | Â | Â | Â | Â | Â | Â |
21F | GAATTTAATCTCTGATTTCTCTAC | 168 | 53 | 60.5 | 56 | 48–57 | 46–55 |
21R | GAGTGAATGTGAATACATTTCC | Â | Â | Â | Â | Â | Â |
22F | AAATAGGTTAAAATAAAGTGTTAT | 206 | 53 | 48 |  | 48–57 |  |
22R | GACTAATTACATACCATTAAG | Â | Â | Â | Â | Â | Â |
23F | CTCTGTATTCACTTTCCATG | 250 | 54 | 57 |  | 52–61 |  |
23R | ACAGTTAGTCACCCTACCCA | Â | Â | Â | Â | Â | Â |
24F | GCTCAGTATAACTGAGGCTG | 249 | 54 | 58.5 |  | 51–60 |  |
24R | CTCTGAGTCAGTTAAACAGT | Â | Â | Â | Â | Â | Â |
25F | AGTGCTGTGGTATGGTTAAG | 323 | 56 | 58 | 56.5 | 55–64 | 54–63 |
25R | TTGCTCTGAAAATTTGGTCATA | Â | Â | Â | Â | Â | Â |
26F | ATCCTGGACTACTGGAAACA | 393 | 53 | 63 | 58.5 | 56–65 | 51–60 |
26R | AGTTAATTCAGGAGGCTTCA | Â | Â | Â | Â | Â | Â |