Oligonucleotide primers, MgCl2 concentrations, and annealing temperatures for FH PCR. DHPLC melting temperatures correspond to column temperatures used in heteroduplex analysis. Mitochondrial targeting signal is defined as exon 0 (FH0)
Exon | Primer | Primer sequence (5‘ > 3‘) | Product size (bp) | MgCl2 (mmol/l) | Annealing temp (°C) | DHPLC melting temp (°C) |
---|---|---|---|---|---|---|
0 | FH0F | TGT GAG GCT GTT GAT TGG AT | 299 | 0 | 59 | 66 |
FH0R | GGA GGG CTG AAG GTC ACT G | |||||
1 | FH1F | AAG ATG CGA TTA CTT TTG ATC C | 256 | 2.8 | 58 | 58 |
FH1R | TGA ATA CAG CCT ACT TCA TCCA A | |||||
2 | FH2F | CTG CCA AAA TAA TAA ACT TCC ATG C | 254 | 1.4 | 58 | 53/56 |
FH2R | GCC AGA GCA TAT CGT CAT CC | |||||
3 | FH3F | TGT GGG TCA ACT GTA TTC AAA C | 321 | 1.4 | 58 | 57 |
FH3R | CAA TCT CAG GTA TGC TTT TCA A | |||||
4 | FH4F | GCT GGG TTT TGA GTA GTT AGT TGG | 349 | 1.4 | 58 | 56/57 |
FH4R | GGC CAT TTG TAC CAA GCT CT | |||||
5 | FH5F | TTTGCTCATCATAAGATTTGAAGT | 381 | 2.8 | 60 | 59 |
FH5R | CAGACCACGTATAATGAGAAATGAA | |||||
6 | FH6F | GTT CAC CCA TCT AGG ATA TTT TTC T | 340 | 2.8 | 60 | 61 |
FH6R | GCT TCG GGA TAA CTT TAA ACA AA | |||||
7 | FH7F | ATGGTTGGGCCTTGCTTTAT | 300 | 2.8 | 58 | 54/57 |
FH7R | CCAAGATAATAAGCCTTTGGTCA | |||||
8 | FH8F | CATGTTGCCTTAGTAATGTCTCTCTC | 317 | 1.4 | 58 | 56 |
FH8R | TGCTGTTCTCAAACACTGATCC | |||||
9 | FH9F | CAATTATGTCACCTTTTGCTTTAGG | 312 | 2.8 | 58 | 53/56 |
FH9R | GCAGTTTCCTTTCAAACTTATCC |