Table 1.
Primer Sequences and DHPLC Conditions
| Name | Primer sequence (5′ to 3′) | PCR size | Tm* PCR (°C) | DHPLC gradient %B | DHPLC† temperatures (°C) |
|---|---|---|---|---|---|
| MLH1 primer list | |||||
| MLH1-1F | aggtgattggctgaaggcac | 231 | 62 | 47–61 | 62, 63 |
| MLH1-1R | gcccgttaagtcgtagccct | ||||
| MLH1-2F | atgtacattagagtagttgcagactgataaatt | 221 | 57 | 46–60 | 56, 57 |
| MLH1-2R | agtttccagaacagagaaaggtcc | ||||
| MLH1-3F | caagaaaatgggaattcaaagagat | 241 | 55 | 47–61 | 55, 56 |
| MLH1-3R | ctaacaaatgacagacaatgtcatcac | ||||
| MLH1-4F | cctttggtgaggtgacagtgg | 221 | 56 | 46–60 | 57, 58, 59 |
| MLH1-4R | caggattactctgagacctaggcaa | ||||
| MLH1-5F | ttttccccttgggattagtatctatc | 227 | 53 | 47–61 | 55, 57 |
| MLH1-5R | ccctgaaaacttagaagcaattttattt | ||||
| MLH1-6F | ggacatcttgggttttattttcaag | 235 | 56 | 47–61 | 57, 58 |
| MLH1-6R | tgttcaatgtatgagcactagaacaca | ||||
| MLH1-78F | gggctctgacatctagtgtgtgtt | 417 | 56 | 52–66 | 56, 57, 58 |
| MLH1-78R | aaaataatgtgatggaatgataaacca | ||||
| MLH1-9F | tctgattcttttgtaatgtttgagttttg | 241 | 55 | 47–61 | 56, 57 |
| MLH1-9R | cataaaattccctgtgggtgtttc | ||||
| MLH1-10F | ctgaggtgatttcatgactttgtgt | 251 | 59 | 48–62 | 58, 59, 60 |
| MLH1-10R | gaggagagcctgatagaacatctgt | ||||
| MLH1-11F | gtgggctttttctccccct | 281 | 58 | 49–63 | 58, 60, 62 |
| MLH1-11R | ctctcacgtctggccgg | ||||
| MLH1-12 new F | ttttttaatacagactttgctaccaggac | 436 | 55 | 54–68 | 59, 60, 61 |
| MLH1-12 new R | gttttattacagaataaaggaggtaggctg | ||||
| MLH1-13F | ccaaaatgcaacccacaaaatt | 282 | 58 | 49–63 | 58, 59, 60 |
| MLH1-13R | aaccttggcagttgaggcc | ||||
| MLH1-14F | ggtgtctctagttctggtgcctg | 271 | 58 | 48–62 | 59, 60 |
| MLH1-14R | tgcctgtgctccctgga | ||||
| MLH1-15F | cccattttgtcccaactggtt | 203 | 57 | 45–59 | 56, 57, 58 |
| MLH1-15R | gagagctactattttcagaaacgatcag | ||||
| MLH1-16F | tgggaattcaggcttcatttg | 292 | 58 | 49–63 | 58, 59, 60 |
| MLH1-16R | gcacccggctggaaatt | ||||
| MLH1-17F | gcactggagaaatgggatttg | 221 | 59 | 46–60 | 58, 59, 60 |
| MLH1-17R | cctccagcacacatgcatg | ||||
| MLH1-18F | agtctgtgatctccgtttagaatgag | 242 | 57 | 47–61 | 56, 58, 59 |
| MLH1-18R | ttgtatgaggtcctgtcctagtcct | ||||
| MLH1-19F | catcagccaggacaccagtg | 288 | 58 | 49–63 | 58, 59, 60 |
| MLH1-19R | cggaatacagagaaagaagaacaca | ||||
| MSH2 primer list | |||||
| MSH2-1-diag F | ttcgacatggcggtgc | 285 | 67 | 48–62 | 66, 67 |
| MSH2-1 new R | gtccctccccagcacg | ||||
| MSH2-2F | gaagtccagctaatacagtgcttga | 301 | 53 | 49–63 | 52, 54, 55 |
| MSH2-2R | aaacacaattaaattcttcacatttttatttt | ||||
| MSH2-3F | agagtttggatttttcctttttgc | 432 | 57 | 52–66 | 56, 57, 58 |
| MSH2-3R | tcatgtcaattaaagagcctttcc | ||||
| MSH2-4F | ttcatttttgcttttcttattcctttt | 316 | 50 | 50–64 | 52, 55, 57 |
| MSH2-4R | atatgacagaaatatccttctaaaaagtcactat | ||||
| MSH2-5F | actggatccagtggtatagaaatcttc | 285 | 53 | 49–63 | 52, 54, 56 |
| MSH2-5R | gcttcttcagtatatgtcaatgaaaaca | ||||
| MSH2-6F | gcgtagtaaggttttcactaatgagc | 251 | 56 | 48–62 | 56, 57, 58 |
| MSH2-6R | catgtgggtaactgcaggttaca | ||||
| MSH2-7F | tgagacttacgtgcttagttgataaattt | 341 | 53 | 50–64 | 52, 55, 56 |
| MSH2-7R | gcacattgccaagtatatattgtatgag | ||||
| MSH2-8F | tgatgcttgtttatctcagtcaaaatt | 275 | 53 | 48–62 | 53, 54, 55 |
| MSH2-8R | aatctacaaactttcttaaagtggcctt | ||||
| MSH2-9 new F | gtctttacccattatttataggattttgtca | 217 | 56 | 46–60 | 56, 57, 58 |
| MSH2-9 new R | gtatagacaaaagaattattccaacctcc | ||||
| MSH2-10F | attgaaaaatggtagtaggtatttatggaa | 274 | 54 | 48–62 | 54, 55, 56 |
| MSH2-10R | cacatcatgttagagcatttaggga | ||||
| MSH2-11F | atatgtttcacgtagtacacattgcttcta | 249 | 54 | 47–61 | 54, 55 |
| MSH2-11R | tcaaatatcatgatttttcttctgttacc | ||||
| MSH2-13R | tcacaggacagagacatacatttctatct | ||||
| MSH2-14F | tgtggcatatccttcccaatg | 452 | 55 | 52–66 | 55, 56, 57 |
| MSH2-14R | aataatttatactaacttagaataaggcaattactgat | ||||
| MSH2-15F | tacataaattgctgtctcttctcatgc | 311 | 57 | 50–64 | 57, 58 |
| MSH2-15R | aaaaaccttcatcttagtgtcctgttt | ||||
| MSH2-16 new F | taattactaatgggacattcacatgtgt | 230 | 55 | 47–61 | 55, 56 |
| MSH2-16 new R | taccttcattccattactgggattt | ||||
| MSH6 primer list | |||||
| MSH6Exon 1F | tgttgattggccactggg | 463 | 66 | 53–58 | 66, 67, 68 |
| MSH6Exon 1R | caaccccctgtgcgagcctc | ||||
| MSH6Exon 2F | taactgcctttaaggaaacttgacca | 330 | 60 | 50–55 | 59, 60, 61 |
| MSH6Exon 2R | tcatatagaaaaaagtctgcctgtctg | ||||
| MSH6Exon 3F | ctggtcttgaactgctgggat | 289 | 58 | 49–54 | 58, 59, 60 |
| MSH6Exon 3R | cccctttcttcccccatc | ||||
| MSH6Exon 4-1F | tgcacgggtaccattataaagtca | 450 | 58 | 52–57 | 58, 59, 60 |
| MSH6Exon 4-1R | gtattcttggtttctgatgaaatgctag | ||||
| MSH6Exon 4-2F | gaaggaaacgccctcagc | 420 | 58 | 52–57 | 58, 59, 60 |
| MSH6Exon 4-2R | cagttgcctttcatgaataccag | ||||
| MSH6Exon 4-3F | ccacatggatgctcttattgga | 420 | 58 | 52–57 | 58, 59, 60 |
| MSH6Exon 4-3R | tcatctgaaaactgacctatgaaaaact | ||||
| MSH6Exon 4-4F | tttgttgatacttcactgggaaagtt | 420 | 57 | 52–57 | 57, 58, 59 |
| MSH6Exon 4-4R | ctcctgatcaataaggcattttttg | ||||
| MSH6Exon 4-5F | ctctaggtggttgtgtcttctacctc | 420 | 57 | 52–57 | 57, 58, 59 |
| MSH6Exon 4-5R | tgagtagcctctcaagatctggaa | ||||
| MSH6Exon 4-6F | cgaagttgtagagcttctaaagaagct | 480 | 57 | 53–58 | 56, 57, 58 |
| MSH6Exon 4-6R | gtcctacagccaattctgttgc | ||||
| MSH6Exon 4-7F | agcctcctggaatacctagagaaac | 420 | 58 | 52–57 | 58, 59, 60 |
| MSH6Exon 4-7R | acttatttttagggataatatacagctggc | ||||
| MSH6Exon 5F | cacttaggctgataaaaccccc | 386 | 57 | 51–56 | 57, 58, 59 |
| MSH6Exon 5R | gtatgttattcctaatgtcacaaatgacttt | ||||
| MSH6Exon 6F | aagacaaaagtttatgaaactgttactacca | 250 | 56 | 48–53 | 56, 57, 59 |
| MSH6Exon 6R | agaagcaaatatcttttatcacatctaaatg | ||||
| MSH6Exon 7F | taacctagaagatgaatttatgtaatatgattt | 224 | 53 | 46–51 | 53, 54, 55 |
| MSH6Exon 7R | ttcagataatcttctataaaaatagttatttgt | ||||
| MSH6Exon 8F | tgagttacttccttatgcatattttact | 275 | 57 | 48–53 | 56, 57, 58 |
| MSH6Exon 8R | aatattagcgatacatgtgctagca | ||||
| MSH6Exon 9F | tgctagcacatgtatcgctaatatt | 320 | 56 | 50–55 | 56, 57, 58 |
| MSH6Exon 9R | gcatcatcccttcccctttta | ||||
| MSH6Exon 10F | gaagggatgatgcactatgaaaaa | 296 | 52 | 49–54 | 52, 56, 57 |
| MSH6Exon 10R | gtagaaggtagataagaattaaaagggtttaattt |
*
Tm for the fragment.
†
Optimum temperatures for DHPLC analysis were empirically determined using the predicted fragment melting profile generated by WAVEMAKER software. (table continued)