Aging Aging

268 Taylor et al.Table 2H-strand PCR Primers Used for Amplification of mtDNAfor Dye Primer SequencingPrimer Position Sequence (5'-3')01R 1190–1172 CAGGAAACAGCTATGACCGATATGAAGCACCGCCAGG02R 1801–1782 CAGGAAACAGCTATGACCTCATCTTTCCCTTGCGGTAC03R 2444–2426 CAGGAAACAGCTATGACCTGAGCATGCCTGTGTTGGG04R 3074–3054 CAGGAAACAGCTATGACCTGAACTCAGATCACGTAGGAC05R 3640–3622 CAGGAAACAGCTATGACCCTAGGCTAGAGGTGGCTAG06R 4239–4219 CAGGAAACAGCTATGACCGATTGTAATGGGTATGGAGAC07R 4866–4848 CAGGAAACAGCTATGACCATGTGAGAAGAAGCAGGCC08R 5570–5551 CAGGAAACAGCTATGACCAGTATTGCAACTTACTGAGG09R 6188–6171 CAGGAAACAGCTATGACCGGGAAACGCCATATCGGG10R 6781–6761 CAGGAAACAGCTATGACCAATATATGGTGTGCTCACACG11R 7398–7379 CAGGAAACAGCTATGACCGGCATCCATATAGTCACTCC12R 8009–7990 CAGGAAACAGCTATGACCCTCGATTGTCAACGTCAAGG13R 8641–8621 CAGGAAACAGCTATGACCTGATGAGATATTTGGAGGTGG14R 9231–9212 CAGGAAACAGCTATGACCGATAGGCATGTGATTGGTGG15R 9867–9848 CAGGAAACAGCTATGACCGGATGAAGCAGATAGTGAGG16R 10516–10497 CAGGAAACAGCTATGACCAGTGAGATGGTAAATGCTAG17R 11032–11013 CAGGAAACAGCTATGACCTCGTGATAGTGGTTCACTGG18R 11708–11689 CAGGAAACAGCTATGACCTTATGAGAATGACTGCGCCG19R 12361–12341 CAGGAAACAGCTATGACCTGGTTATAGTAGTGTGCATGG20R 13005–12987 CAGGAAACAGCTATGACCTTTGCCTGCTGCTGCTAGG21R 13614–13595 CAGGAAACAGCTATGACCTATTCGAGTGCTATAGGCGC22R 14276–14258 CAGGAAACAGCTATGACCGGTTGATTCGGGAGGATCC23R 14928–14911 CAGGAAACAGCTATGACCGTTGAGGCGTCTGGTGAG24R 15419–15400 CAGGAAACAGCTATGACCTGTAGTAAGGGTGGAAGGTG25R 16067–16048 CAGGAAACAGCTATGACCGTCAATACTTGGGTGGTACCD01R 16545–16526 CAGGAAACAGCTATGACCAACGTGTGGGCTATTTAGGCD02R 559–542 CAGGAAACAGCTATGACCGGGTTTGGTTGGTTCGGGD03R 803–786 CAGGAAACAGCTATGACCGGTGTGGCTAGGCTAAGCto give consistently high quality sequence data. QIAquick PCR Purificationcolumns (Qiagen) are both quick and easy to use and have the advantage thatthe final elution volume can be varied to give a template concentration appropriatefor the length of fragment to be sequenced.1.2. Last Hot Cycle PCRThe demonstration of mtDNA heteroplasmy associated with a biochemicaldefect remains the most compelling piece of evidence of pathogenicity for anymutation as it implies a recent mutational event. In view of the high levels of