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DNA microarray for rapid detection of mitochondrial DNA haplotypes of chum salmon
被引:12
作者:
Moriya, S
Urawa, S
Suzuki, O
Urano, A
Abe, S
机构:
[1] Nisshinbo Ind Inc, Ctr Res & Dev, Midori Ku, Chiba 2670056, Japan
[2] Natl Salmon Resources Ctr, Genet Sect, Toyohira Ku, Sapporo, Hokkaido 0620922, Japan
[3] Hokkaido Univ, Div Biol Sci, Grad Sch Sci, Sapporo, Hokkaido 0600810, Japan
[4] Hokkaido Univ, Lab Anim Cytogenet, Ctr Adv Sci & Technol, Kita Ku, Sapporo, Hokkaido 0600810, Japan
[5] Hokkaido Univ, Lab Breeding Sci, Div Marine Biosci, Grad Sch Fisheries Sci, Hakodate, Hokkaido 0418611, Japan
关键词:
DNA microarray;
chum salmon;
mtDNA;
haplotype;
stock identification;
D O I:
10.1007/s10126-004-1100-2
中图分类号:
Q81 [生物工程学(生物技术)];
Q93 [微生物学];
学科分类号:
071005 ;
0836 ;
090102 ;
100705 ;
摘要:
For use in genetic stock identification, we developed an oligonucleotide (DNA) microarray hybridization method for rapid and accurate detection of nucleotide sequence variations in 20 previously identified variable nucleotide sites in about 500 bp within the 5' half of the control region of mitochondrial DNA of chum salmon (Oncorhynchus keta). The method includes immobilization of synthesized oligonucleotides containing respective polymorphic sites on a glass slide precoated with polycarbodiimide resin, a 2-hour hybridization with DNA microarray of biotinylated polymerase chain reaction fragments spanning the 5' variable portion followed by short washing, and visualization of hybridization signals by conventional ABC method and scanner-assisted computation of signal intensity on a computer. The entire process of hybridization and detection was completed within 4 hours. The resulting DNA microarray could detect all of the single nucleotide mutations and therefore could be used to identity the sequence variations defining 30 mtDNA haplotypes of chum salmon as revealed previously by nucleotide sequence analysis.
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页码:430 / 434
页数:5
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