We performed BLAST searches of our ND2
and aconitase sequences against the GenBank nucleotide database to identify candidate parent species with high percent sequence identity.
We gathered ND2
sequences from five members of the E.
Sample numbers followed by an asterisk identify samples used in all analyses; remaining specimens were sequenced only for ND2
For the mitochondrial ND2
sequences derived from ancient DNA sources, we performed alignments of each individual fragment against homologous sequences from fresh samples as well as BLAST searches in order to verify for erroneous base calls or amplification of pseudogenes (Sorenson and Quinn 1998).
Three target regions of the arowana mitochondrial DNA were selected for sequencing: 1044bp encompassing the complete ND2
, 628 bp of cytb gene region and 72 bp of tRNA-Thr.
Correct is 12 Nd2
Bxe2 13 Qxe2 Nh5 14 Be3 Nd7 15 a5 or 15 g4.
change Gene B1T 5 G3010A RNR2 B1T 7 A4769G ND2
B1T 11 A9448G CO3 B1T 16 A15326G Cyt-b S2T 3 C476T RNR1 S2T 8 G5979A CO1 S2T 10 C7956T CO2 S2T 12 C10527T ND4L S2T 13 C12465T ND5 S2T 1 C16520T D-loop B8T 2 A263G R sequence B8T 14 A13032G ND5 B8T 16 C15409T Cyt-b B8T 17 A16051G D-loop S3T 6 C3992T ND1 S3T 7 T5004C ND2
S3T 10 G8269A CO2 S3T 11 G9123A ATP6 S3T 12 A10044G tG S3T 16 A15326G Cyt-b Detected in Detected in Homoplasmy or Tumor uniplex?
is a well-known mtDNA marker in birds and has been shown to be particularly informative and approximately neutrally evolving (Zink et al.
Image Format: JP2, JPG, TIFF, BMP, GIF, PNG, ND2
, JFF, JTF, AVI, ICS/IDS
WHITE:COLIN SEARLE BLACK: ALASTAIR DAWSON Queen's Pawn1 d4 Nf6 2 Nf3 d6 3 Bf4 g6 4 e3 Bg7 5 Be2 0-0 6 h3 Nbd7 7 0-0 Re8 8 Re1 e5 9 dxe5 dxe5 10 Nxe5 Nh5 11 Bxh5 Nxe5 12 Be2 Qe7 13 c3 Be6 14 Nd2
Rad8 White already lacks space.
PCR products of mitochondrial genes (16S, ND2
, ATPase6, and ND5) and the nuclear ATPsyn[beta] gene (Table 2) were subcloned into the plasmid vector pGEM[R]-T Easy (pGEM-T and pGEM-T Easy Vector Systems; Promega) according to the manufacturer's instructions.
Primers H5578 (5' CCT TGA AGC ACT TCT GGG AAT CAG A3') and L5215 (5' TAT CGG GCC CAT ACC CCG AAA AT 3'; Hackett 1996) were used to amplify 360 bp of the ND2
gene of all specimens.