Universal nucleic acid probe set and method for utilization...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/09 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2735250

A nucleic acid probe set includes (A) a fluorescent probe and (B) a binding probe. The fluorescent probe (A) is formed of an oligonucleotide, which includes (a) a nucleotide unit labeled with (d) a fluorescent substance. The binding probe (B) is formed of an oligonucleotide having (b1) a fluorescent probe binding region, which can hybridize to the fluorescent probe (A), and (b2) a target nucleic acid binding region, which can hybridize to a target nucleic acid sequence (C). The fluorescent substance (d) is a fluorescent substance which changes in fluorescent character upon interaction with guanine. At least one of nucleotide units which constitute the fluorescent probe (A) is an artificial nucleotide unit having a function to raise a dissociation temperature between the probe (A) and the fluorescent probe binding region (b1). The nucleic acid probe is provided with an improved fluorescence quenching efficiency.

L'invention porte sur un ensemble de sondes d'acides nucléiques caractérisé par le fait qu'il comprend : une sonde fluorescente (A) qui comprend un oligonucléotide comprenant un nucléotide (a) marqué par une substance fluorescente (d); et une sonde de liaison (B) qui comprend un oligonucléotide ayant une région de liaison à la sonde fluorescente (b1) capable de s'hybrider avec la sonde fluorescente (A) et une région (b2) de liaison à l'acide nucléique cible capable de s'hybrider avec une séquence d'acide nucléique cible (C), la substance fluorescente (d) pouvant changer son caractère fluorescent lorsque la substance fluorescente (d) interagit avec la guanine, et au moins l'un des nucléotides constituant la sonde fluorescente (A) étant un nucléotide artificiel capable d'augmenter la température de fusion entre la sonde (A) et la région de liaison à la sonde fluorescente (b1). L'ensemble de sondes d'acides nucléiques est amélioré en ce qui concerne la vitesse d'extinction de fluorescence et présente à la fois une sonde fluorescente et une sonde de liaison.

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