Primer sequences, annealing temperatures, and buffers used for PCR reactions
| Exon | Forward primer | Reverse primer | Length | Ta | Buffer | Sequence primer | Orientation | ||
|---|---|---|---|---|---|---|---|---|---|
| M13F, TGTAAAACGACGGCCAGT; M13FX, GTGGTGTAAAACGACGGCCAGT; M13R, CAGGAAACAGCTATGACC; M13RX, GTGGCAGGAAACAGCTATGACC | |||||||||
| * PCR product sequenced in both directions for single coverage | |||||||||
| 1 | 1F.A | AGACCCTCTCCTTGCCTCTT | 1R.0 | M13RX-ATGGAGGGTCGGAGGCTG | 458 | 55.0 | D | M13R | Reverse |
| 2 | 2F.1 | TGGCAAGTAAGTTATTTATGGTC | 2R.1 | M13F-AATCAAAAAGAAAAGAAAGCAA | 288 | 55.0 | B | M13F | Reverse |
| 3 | 3F.1 | GTTTGCCTTAGACTTTAGTTTTT | 3R.1 | M13F-CATCTGTACTTTGGGACATAA | 368 | 55.0 | B | M13F | Reverse |
| 4a | 4aF.0 | TTTGAAAATTTTCATAATAGAAAATGT | 4aR.0 | M13RX-GGTCAAAGCTGCTGTGAG | 435 | 55.0 | C | M13R | Reverse |
| 4b | 4bF.0 | M13FX-TCCTGGCCTCAAGTGGTC | 4bR.0 | M13RX-TTATAAAANCCAGATTGGTGTTC | 386 | 57.0 | B | M13F | Forward |
| 4c | 4cF.0 | M13FX-TCCTAGCAGACAACTATCGA | 4cR.0 | AAAAAAAAATCAATCGTATCCTTA | 582 | 55.0 | C | M13F | Forward |
| 6 | 6F.0 | M13FX-CATGTTTATCTTTTAAAAATGTTGCC | 6R.0 | ATAATGGAAATAATTTTGCCCTCC | 323 | 55.0 | C | M13F | Forward |
| 7 | 7F.0 | M13FX-TGCTATAATATTAGCTACATCTGG | 7R.0 | CCTATGAACTTATCAACGAAGAG | 395 | 55.0 | B | M13F | Forward |
| 8 | 8F.0 | M13FX-TTGTGTGGGTAATGTGTTGA | 8R.0 | AAATATAGTTAGATAAAAACCAATG | 270 | 55.0 | B | M13F | Forward |
| 9 | 9F.1 | TATTTGCTGTTCTTTTTGG | 9R.1 | M13F-AATTTAGCAATACCTTTTGG | 204 | 55.0 | B | M13F | Reverse |
| 9a | 9aF.0 | TCCGCTGTGGCTCAGAACAC | 9aR.0 | M13RX-AGTAGAAGAGGATGCACAGCC | 315 | 60.0 | B | M13R | Reverse |
| 10a | 10aF.0 | M13FX-GATAAACAGAGCATACAACTCA | 10aR.0 | AAATGCAATAGAAAGGAGGTGA | 270 | 55.0 | B | M13F | Forward |
| 10b | 10bF.A | AACTATTGAGTGTTCTACTATACC | 10bR.0 | M13RX-TTGGCGATTCAGCTAAACC | 374 | 55.0 | G | M13R | Reverse |
| 10c | 10cF.1 | M13F-CGTCCAGCCTAGTTCTAGAA | 10cR.1 | ACCCCTTCTTTCTCCAT | 444 | 55.0 | C | M13F | Forward |
| 11 | 11F.0 | M13FX-ATAAGTACTCCAGTGTTATGT | 11R.0 | TAAAGTTGAAATTTAAAAAATTAAAGTAC | 179 | 52.0 | C | M13F | Forward |
| 12a | 12aF.0 | M13FX-AAACCTTACAAGAAAAACTAAGCT | 12aR.0 | ATTACCATTCCAAATATTCTTCCA | 325 | 55.0 | C | M13F | Forward |
| 12b | 12bF.0 | M13FX-TTTCTAGTAAATCTCCTTCAAGT | 12bR.0 | ATGAAATTTACCAAATTTCATTCAG | 404 | 55.0 | C | M13F | Forward |
| 13 | 13F.1 | AAGATATTTTGGGGTTTGAA | 13R.1 | M13R-GCCATGTGCTTTGAGG | 508 | 55.0 | B | M13R | Reverse |
| 14 | 14F.0 | M13FX-AGCTTATCAGGTTCTCCATTGG | 15R.0 | M13R-AGTTAACAGACAAAAGTCAACTTTACAG | 535 | 55.0 | D | M13F | Forward |
| 15 | 14F.0 | M13FX-AGCTTATCAGGTTCTCCATTGG | 15R.0 | M13R-AGTTAACAGACAAAAGTCAACTTTACAG | 535 | 55.0 | D | M13R | Reverse |
| 16 | 16F.0 | M13FX-TGGATAAAGCATAATTTGTCAAGT | 16R.0 | TAGAGAAAGGTGAAAAATAAGAG | 571 | 55.0 | B | M13F | Forward |
| 17 | 17F.0 | CTGTGTGTTTAGATCAGTCA | 17R.0 | M13RX-TTTATCAATTACTACCAGTATCAG | 339 | 55.0 | B | M13R | Reverse |
| 18 | 18F.0 | AGAAGTTGTGTACGTTCTTTTCT | 18R.0 | M13RX-TCCTTTCTACCAATAACCGC | 388 | 55.0 | I | M13R | Reverse |
| 19a | 19aF.0 | M13FX-ATGTCACTTAGGTTATCTGG | 19aR.0 | TGTAATTAAGTAGTTATAACTCTC | 290 | 60.0 | B | M13F | Forward |
| 19b | 19bF.1 | M13F-ATTTGAGGGGAAGTGAA | 19bR.2 | M13R-ATGGGTATTATTTCTTTGCT | 312 | 55.0 | E | M13R | Reverse |
| 20 | 20F.0 | M13FX-CATTCACACCATGCACATATGATTG | 21R.0 | M13R-GATTTGCTATGTGCCAGGCAC | 703 | 55.0 | E | M13F | Forward |
| 21 | 20F.0 | M13FX-CATTCACACCATGCACATATGATTG | 21R.0 | M13R-GATTTGCTATGTGCCAGGCAC | 703 | 55.0 | E | M13R | Reverse |
| 22 | 22F.0 | M13FX-TGCTACTCTTTAGCTTCCTAC | 22R.0 | ACAGCGGTTCTATGTGAAAAG | 537 | 55.0 | B | M13F | Forward |
| 23-2 | 23-2F.0 | M13FX-CTTAATGTCTGTATAAGAGTCTC | 23-2R.0 | ACTTTAGATTAATAATGGTAATCTC | 290 | 55.0 | C | M13F | Forward |
| 23a | 23aF.0 | CAGAAATAGTATACATGATTGGGT | 23aR.0 | M13RX-CTATTTTCTGCCAGAATTAGTAGA | 465 | 55.0 | C | M13R | Reverse |
| 24 | 24F.0 | M13FX-TTGAACTCTTTTGTTTTCATGTCTT | 24R.0 | GGAATTTAAGATAGCTAGATTATC | 289 | 55.0 | C | M13F | Forward |
| 25 | 25F.0 | M13FX-AATATAATAATTATATTTGGGAAGGT | 25R.0 | GAAAATATTTGATTCAAACAGAGC | 360 | 55.0 | B | M13F | Forward |
| 26 | 26F.1 | M13F-CCCTCCATATTTGTAATCTT | 26R.1 | TTAAACGGAGAGTGTTCAC | 324 | 58.0 | B | M13F | Forward |
| 27a | 27aF.1 | AGGGCATTTTAATCTTTTAT | 27aR.1 | M13F-GCAAACTCTCCTTCTCAAC | 406 | 55.0 | F | M13F | Reverse |
| 27b | 27bF.1 | M13R-GCCAAATTACCCTTTAGAATG | 27bR.1 | AACCAAACTTGCCATCTCT | 348 | 55.0 | C | M13R | Forward |
| 28F* | 28F.1 | M13F-GACTTTGAAGAATTGTTTTATA | 28R.0 | M13RX-CTATAGGGGTAGGACACC | 693 | 55.0 | E | M13F | Forward |
| 28R* | 28F.1 | M13F-GACTTTGAAGAATTGTTTTATA | 28R.0 | M13RX-CTATAGGGGTAGGACACC | 693 | 55.0 | E | M13R | Reverse |
| 29F* | 29F.1 | M13F-TACAATGGTGGGAACTCTTCCTTA | 29R.0 | M13RX-ACCAACACTGCATACCTTCCAATT | 635 | 55.0 | E | M13F | Forward |
| 29R* | 29F.1 | M13F-TACAATGGTGGGAACTCTTCCTTA | 29R.0 | M13RX-ACCAACACTGCATACCTTCCAATT | 635 | 55.0 | E | M13R | Reverse |
| 30 | 30F.0 | GCCTCACAGTGCTCTTATGG | 30R.0 | M13RX-CCTTCAGAGTTCCTTCAGTTAC | 628 | 55.0 | C | M13R | Reverse |
| 31 | 31F.0 | TGTTGATGTGATTTTCATTGACCA | 31R.0 | M13RX-CCAATGTGGCACCAGATAAATAT | 337 | 55.0 | B | M13R | Reverse |
| 32 | 32F.0 | M13FX-ATCTAGTATTTTTGAGGCCTCAG | 32R.0 | CAGATATGCTATAGTACAGAAGG | 334 | 55.0 | C | M13F | Forward |
| 33 | 33F.0 | M13FX-TATCTGTTTTATCATCAGGAGG | 33R.1 | M13R-TAAAATGGAGAAAGGAACTGG | 497 | 55.0 | E | M13R | Reverse |
| 34 | 34F.1 | M13F-AAAATGAAACATGGAACTTTAG | 34R.0 | M13RX-TAAGCATTAAGTACAAAATAGCAC | 439 | 55.0 | E | M13F | Forward |
| 35 | 35F.0 | ATGTATTCAGAGTATCCCCTT | 35R.0 | M13RX-ATTTAAACGTCTTTTAGAAAAATG | 458 | 55.0 | C | M13R | Reverse |
| 37 | 37F.0 | M13FX-ATTCCGAGATTCAGTTTAGGAG | 37R.0 | AGTAACATTCAACACTGATACCC | 256 | 55.0 | H | M13F | Forward |
| 38 | 38F.1 | ACATGGGTAATTTAGGAAGATAAG | 38R.1 | M13F-CAACAAGAAAAGATGGAAGA | 335 | 55.0 | C | M13F | Reverse |
| 39 | 39F.0 | CTACTGTGTGAACCTCATCAACC | 39R.0 | M13RX-GTAAGACATAAGGGCTAACTTACTTC | 306 | 55.0 | I | M13R | Reverse |
| 40 | 40F.0 | GGGAAGAAGACCTCAGCAGAT | 40R.0 | M13RX-AACTTTCTGCTCTGCCACGCA | 345 | 55.0 | I | M13R | Reverse |
| 41 | 41F.0 | TTCATCCTGTTTTAAGTCACACTTG | 41R.0 | M13RX-TTGCCTCCATTAGTTGGAAAATTG | 295 | 55.0 | H | M13R | Reverse |
| 42 | 42F.0 | GAAGGAGCAAACGATGGTTG | 42R.0 | M13RX-AAACTTTGCTACACTGACATGG | 371 | 55.0 | C | M13R | Reverse |
| 43 | 43F.0 | GTATTAGAGCTTTCTTTGAGTC | 43R.0 | M13RX-GAATGGTAGAGTTTATCTTTTG | 421 | 55.0 | C | M13R | Reverse |
| 44 | 44F.2 | M13R-GGTGAAGTGATTATCCAGGTG | 44R.2 | TGTTATCTGTTTGGGTGAGAAG | 328 | 55.0 | E | M13R | Forward |
| 45 | 45F.0 | M13R-AGATAACAATTCAGCCACAAAG | 45R.0 | TAAAGACAGGCACGAAGGT | 225 | 55.0 | F | M13R | Forward |
| 46 | 46F.1 | M13R-ATTTTGGCACATTATTCTGG | 46R.1 | GCGCATGTTAGCAAGTT | 316 | 55.0 | C | M13R | Forward |
| 47 | 47F.0 | CTGTTACAATTAAAAGATACCTTGC | 47R.0 | M13RX-GTGTTCTTAAAGCAGGCATAC | 204 | 55.0 | H | M13R | Reverse |
| 48 | 48F.0 | TTTTGGCTTCAGATGGGGATTTAC | 48R.0 | M13RX-AAGGGAATTCCTAATGTTGGTGTC | 373 | 55.0 | B | M13R | Reverse |
| 48a | 48aF.0 | ATCTAGTATCTAATTGTATTTCACCC | 48aR.0 | M13RX-AGACTGAGCTTACAGGGAC | 265 | 55.0 | A | M13R | Reverse |
| 49 | 49F.0 | M13FX-CTGGGAGAAACAGGCTATAC | 49R.0 | CACTTTCTTTGCAGTTGTTCTG | 639 | 58.0 | E | M13F | Forward |