Table 1. Overview of multiplex system for mouse mtDNA genome sequencing.
Eight tubes (A–H) per sample were used to amplify the entire murine mitochondrial genome. Two PCR products (1, 2) were generated in each tube. These pairs of PCR products were subsequently digested with various restriction enzymes (RE) to generate restriction fragments (RF) and single nucleotide changes were detected using TGCE. The Start, End and Size of each PCR product is listed along with the primers used to generate the PCR products, the restriction enzymes used and the sizes of the fragments generated via the restriction enzymes.
| Tube ID |
Start | End | Size | Forward Primer | Reverse Primer | Restriction Enzyme |
Restriction Fragment |
|---|---|---|---|---|---|---|---|
| A1 | 1050 | 2340 | 1291 | TCTGGCCTACACCCAGAAGATTTCA | CTGGGTCAATAAGATATGTTGATTTTACTTTGACT | EcoRI | 759,700,591 |
| A2 | 8465 | 9223 | 759 | TATTAATAAATATTAGCCCACCAACAGCTACCATT | AGTCCATGGAATCCAGTAGCCATGA | ||
| B1 | 7168 | 8461 | 1294 | TACAAGCACAATAGATGCACAAGAAGTTGA | GAGTAGCTCCTCCGATTAGGTGTATTAATAAGTGT | TaqI | 794,597,500,431,338 |
| B2 | 14915 | 16280 | 1365 | TAATCCACTAAACACCCCACCCCATATT | GCGTAATAGAGTATGATTAGAGTTTTGGTTCACG | ||
| C1 | 11399 | 12666 | 1267 | CACCCAACGCGGCAAACTAAC | CTAGTTGGCTTGATGTAGAGAAGGCAATG | TaqI | 770,681,587,487 |
| C2 | 16159 | 1121 | 1257 | TCAAACCCTATGTCCTGATCAATTCTAGTAGTTC | TTTGTGTAGGGCTAGGGCTAGGATTAGTT | ||
| D1 | 5828 | 6299 | 472 | TATTATCAACATGAAACCCCCAGCCATA | AAGTCAGCTAAATACTTTGACACCGGTAGG | DraI | 694,595,472 |
| D2 | 9220 | 10508 | 1289 | GACTCCATGTAATTATTGGATCAACATTCCT | AATTAGTTCAGTTGCTGAAAAGGTTATGATTAGG | ||
| E1 | 2248 | 3507 | 1260 | GGTTGGGGTGACCTCGGAGAAT | CTGGTAAATTGATATAGTATAGGGGTCCTAGGAAGA | DraI | 793,701,467 |
| E2 | 6550 | 7250 | 701 | CATGAGCAAAAGCCCACTTCG | AGGGGAGAGCAATTATGATAAGGATTACAGC | ||
| F1 | 6123 | 6700 | 578 | GAACCTTTCGGCTATATAGGAATAGTATGAGCAAT | GATCCTATAGAAGAGACAGTGTTTCATGTGGTG | DraI | 801,578,468 |
| F2 | 13701 | 14969 | 1269 | CTCCCAAACCATCAAGATTAATTACTCCAA | TGCAAATAGGAAATATCATTCGGGTTTAAT | ||
| G1 | 4661 | 5988 | 1328 | TCCCTAGGAGGCCTTCCACCAC | CAAAGAAAGTTGTGTTTAGGTTGCGGTCT | DraI | 809,715,613,501 |
| G2 | 10194 | 11503 | 1310 | CTACCACTAACCTGACTATCAAGCCCTAAAA | GTTAGAAGAATAAGTGGAATTATGTGAAGGGCTAT | ||
| H1 | 3407 | 4700 | 1294 | ATTCGCGTTATTCTTTATAGCAGAGTACACTAACA | TTGGTAAGAATCCTGTTAGTGGTGGAAG | DraI | 807,703,633,487 |
| H2 | 12487 | 13822 | 1336 | CAATAGTAGTTGCAGGAATTTTCCTACTGGTC | TTGGGGATCTAACTGATTAATTTTGGGTTT | ||