[A new approach to detect a particular DNA sequence by UV-immobilization of its hybridization complex with a highly specific probe resulting from ligation of a tandem of short oligonucleotides in solution].
A new approach was proposed for detecting amplified DNA fragments by hybridization with a highly selective oligonucleotide probe obtained by ligation of a tandem of three short oligonucleotides (pN8 + pN4 + pN8' Bio) in solution, with subsequent UV-immobilization of the hybridization product on a nylon membrane and its colorimetric detection with the streptavidin-alkaline phosphatase technique. Owing to the high selectivity of ligation, the 20-mer ligation product was detected on a membrane only when it was completely complementary to a template fragment. The results showed that any single-nucleotide substitution in the tetramer-binding site can be localized and identified with the use of all 12 possible tetramers.
Kabilov
M R
MR
Institute of Cytology and Genetics, Siberian Division, Russian Academy of Sciences, Novosibirsk, 630090 Russia.
Pyshnyĭ
D V
DV
Dymshits
G M
GM
Gashnikova
N M
NM
Pokrovskiĭ
A G
AG
Zarytova
V F
VF
Ivanova
E M
EM
D000469:Alkaline Phosphatase / Q000737:chemistry; D011088:DNA Ligases / Q000737:chemistry; D015342:DNA Probes; D017403:In Situ Hybridization / Q000379:methods*; D008567:Membranes, Artificial; D025202:Molecular Diagnostic Techniques; D017422:Sequence Analysis, DNA / Q000379:methods*; D012996:Solutions; D019809:Streptavidin / Q000737:chemistry; D020080:Tandem Repeat Sequences; D014466:Ultraviolet Rays
D015342:DNA Probes; D008567:Membranes, Artificial; D012996:Solutions; D019809:Streptavidin; D000469:Alkaline Phosphatase; D011088:DNA Ligases
Novyĭ podkhod k vyiavleniiu analiziruemoĭ posledovatel'nosti DNK putem UF-immobilizatsii ee gibridizatsionnogo kompleksa s vysokospetsifichnym zondom, obrazuiushchimsia pri ligirovanii tandema korotkikh oligonukleotidov v rastvore.