Table 1.
MYH7 primers, amplicon sizes, and DHPLC analysis conditions
| Exon | Forward primer 5′→3′ | Reverse primer 5′→3′ | Amplicon size (bp) | Temp (°C) | % Buffer B* |
| 3 | CCT CTT GAC TCT TGA GCA TGG | GTA CCC CTC TCT GTC ACC CA | 392 | 61.9 | 58.9–68.9 |
| 4 | TCC CTG TGA GAT CCT GGT TC | CAA GGA TGT TGG GAC GAG TT | 398 | 62.5 | 59.0–69.0 |
| 5 | AAC TCG TCC CAA CAT CCT TG | AGG TTA GGA GCT GCA CAG GA | 443 | 61.5 | 59.8–69.8 |
| 6 | ACC TTT CCC CCC CAC CCT CT | GGG CTG GAG GCT GGG ATC A | 114 | 62.0 | 43.0–53.0 |
| 7 | GGT CTC CAG TAG TAT TGT TCA | CAA GAA GGA GGC AGG TGA GAG | 229 | 61.8 | 54.2–64.2 |
| 8 | AGG GAG AAG AGC TCT CAC CTG | AAG TCC CAA GGC CAA GGT CAG | 167 | 63.9 | 50.7–60.7 |
| 9 | CCT TGG GAC TTG GAC TGG TG | AAA GAG GAG AAA AAC AGA GGG AGG | 252 | 59.7 | 55.1–65.1 |
| 10 | CCT TCT TCT CCC CAC CTG TTC | GTC TCA GTC GGT GGC TCT GAC | 150 | 58.4 | 49.4–59.4 |
| 11 | GCG AGC AGC CTC CAT GAG | CCC CTC ACT GCC AAT CCT C | 216 | 62.0 | 53.6–63.6 |
| 12 | CAT CAT ACT TCT TTT TTG GGG TCC | GCC CTC CAT GAC TTG ACA GC | 232 | 64.0, 61.0 | 50.8–60.8 |
| 13 | TTA CAG GCA TGA ACC ACA CAC C | GTG AAC TTG AAA ACT CTC ATC CC | 267 | 62.1 | 55.7–65.7 |
| 14 | AGA TGA TAA TGG GTG GGC AG | GAA ATA GCT GTT GAA TGT GGG AG | 328 | 61.5 | 57.5–67.5 |
| 15 | ACT CAC ACC CAC TTT CTG ACT | GAG GGG CTG CTA TTT TGT CTA | 293 | 61.5 | 56.6–66.5 |
| 16 | TTG ACC ATA GAG CAG AAT CCA TGT | GAC ATT GAA GTG GTG GGG TGT AG | 475 | 61.0 | 60.2–70.2 |
| 17 | TTC TTT TGT TGA CTC TCC TTC | GAT GGG GAG CCA AGT TGG CTG | 134 | 61.7 | 48.0–58.0 |
| 18 | TCT CTA TTG CAC TTT TTG GCC | TGG GTT GGC CTG AGT TTG TGG | 140 | 60.6 | 48.6–58.6 |
| 19 | CAG TCC AGT TCT CAC AGA CTC C | GGC TCC CCC TGT TCT ATG AG | 206 | 62.0 | 53.1–63.1 |
| 20 | GGA TCT GCA GGT GAC CCT GAA T | ACA ACA GGA AAA GCA TCA GAG G | 283 | 59.6 | 56.2–66.2 |
| 21 | ACT CCC CTC ATC CCA GCT CCA | CTT TTT TTC CTG ACA CTG CCC | 342 | 61.2 | 57.8–67.8 |
| 22 | AGG CTC AGC ACT CCT TTC AAT | AAG GCA GAG CAG GGT GGA AGA | 335 | 61.3 | 59.7–69.7 |
| 23 | CCC TCC TAT TTG AGT GAT GTG C | GGT CAG TAT GGT CTG AGA GTC C | 404 | 61.0, 59.1 | 59.1–69.1 |
| 24 | CCA CAG ATG GCA CCA AGC TG | TAT CTA GGC CCC ACA ACT CTC A | 271 | 61.3 | 55.8–65.8 |
| 25 | TGG GCC TGG GCT CCT TCT C | GGC AGC AGG GAG GGG ACA | 197 | 62.9 | 52.6–62.6 |
| 26† | CGC CCG CCG CCG CCC TAC CTC ACC ATC CCT CCT TCC | AGC CCA GGG ACT CAG CAT C | 160 | 63.4 | 52.7–62.7 |
| 27 | GCG TGG GTC TGA GCC TTG TGT | GGG GAG GTG GGA GGA GGA AG | 468 | 65.1, 62.2 | 60.1–70.1 |
| 28 | TGC ACC TCT TAC ACC CCT TCA | GGG GAG ACT GTG GTG GGA AC | 242 | 63.1, 61.1 | 54.7–64.7 |
| 29 | GAG GAG GTG GGG ATA GAG AGG A | GTC AGT GTG CTC CTT GCT TGG | 215 | 63.9 | 53.5–63.5 |
| 30 | AGA AAG CTG AAC CCA CCT CCT | CCA GAA GTC AGG CTG CTC AGA | 264 | 66.5, 64 | 55.6–65.6 |
| 31 | CAT CCT CCC CAC CCT CTG C | GCT CTG GCC TCT CAC TGA ACC | 293 | 63.8 | 56.5–66.5 |
| 32 | CTT GGG GGC TGA AGA GTG AG | CCC CTC CCC AGC CTC TTG | 234 | 62.7 | 54.4–64.4 |
| 33 | TCA AAC CGA GTT ACC GTG TTC | TGA GAA CAG GGA CCA AAA GC | 219 | 62.7 | 53.7–63.7 |
| 34 | CCC TGA CTG TCT GCC TGC ATC | GGG GCA GGA GGA ATC TGG TG | 436 | 62.8 | 59.7–69.7 |
| 35 | GCT CAT GCC CAC TCT CCT GAT | TCA GGA ATC AGC AGG GGA GC | 256 | 63.5 | 55.3–65.3 |
| 36 | TCT ACC CAA CCC TCC CCC AAC | GGA TCG GGT CGG TGG AGT G | 180 | 63.4, 61.4 | 48.6–58.6 |
| 37 | CAG ACC ATG TGC CAC CTC TCT | GCA AAC TCT TCA TTC TCC TCA GC | 388 | 64.9, 62.7 | 58.8–68.8 |
| 38 | CAC CCC CTG CCT ACC CTC TGG | GGG AGG TGG GAG CAT GAG GTG | 232 | 63.9 | 54.3–64.3 |
| 39 | CTC ACC TCA TGC TCC CAC CT | TGA CTA GCA AAG CCC AAA AGAG | 192 | 64.1 | 52.3–62.3 |
| 40 | GAT CCT GGC TTT GTT TCC TTT CA | TGG GGC TTT GCT GGC ACC T | 95 | 60.4 | 41.6–51.6 |
The amplicon size includes the corresponding exon and intron–exon boundaries.
Temperature refers to the oven temperature(s) at which DHPLC analysis was performed.
*Range of elution buffer B that was determined by Navigator software based on amplicon sizes; †the underlining highlights the additional GC clamp.
DHPLC, denaturing high performance liquid chromatography.