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Sequence-specific nucleic acid purification method manner

外国特許コード F110005606
整理番号 RX02P38WO
掲載日 2011年9月9日
出願国 アメリカ合衆国
出願番号 93747109
公報番号 20110034683
公報番号 7972792
出願日 平成21年4月16日(2009.4.16)
公報発行日 平成23年2月10日(2011.2.10)
公報発行日 平成23年7月5日(2011.7.5)
国際出願番号 JP2009001747
国際公開番号 WO2009128266
国際出願日 平成21年4月16日(2009.4.16)
国際公開日 平成21年10月22日(2009.10.22)
優先権データ
  • 特願2008-106642 (2008.4.16) JP
  • 2009WO-JP01747 (2009.4.16) WO
発明の名称 (英語) Sequence-specific nucleic acid purification method manner
発明の概要(英語) (US7972792)
Disclosed is a method for the purification and collection of a nucleic acid comprising a specific nucleotide sequence, which can be carried out within an extremely short period and can achieve both high sequence-specificity and a high collection rate.
Specifically disclosed is a method for the purification of a target nucleic acid comprising a specific nucleotide sequence and contained in a nucleic acid mixture.
The method comprises the steps of: hybridizing a photo-ligating nucleic acid having a group represented by formula (I) as abase moiety with the target nucleic acid to form a hybrid; irradiating the hybrid of the photo-ligating nucleic acid and the target nucleic acid with light to cause the photo-ligation of the hybrid; removing any un-photo-ligated nucleic acid by washing; and irradiating the hybrid of the photo-ligating nucleic acid and the target nucleic acid with light to cause the photo-cleavage of the hybrid.
特許請求の範囲(英語) [claim1]
1. A method of purifying a target nucleic acid having a particular base sequence that is contained in a nucleic acid mixture, the method comprising: hybridizing a photo-ligating nucleic acid (wherein the nucleic acid includes a nucleic acid and a peptide nucleic acid) having a group represented by the following formula (I) as a base moiety:
(wherein in the formula (I), Ra represents a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group or hydrogen; and
R1 and R2 each independently represent a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group or hydrogen),
with the target nucleic acid having a particular base sequence that is contained in a nucleic acid mixture;
irradiating the hybrid of the photo-ligating nucleic acid and target nucleic acid with light to induce photo-ligation of the hybrid;
removing any un-photo-ligated nucleic acid by washing; and
irradiating the hybrid of the photo-ligating nucleic acid and the target nucleic acid with light to induce photo-cleavage of the hybrid.
[claim2]
2. The method according to claim 1, wherein the step of removing any un-photo-ligated nucleic acid by washing is carried out by washing under the washing conditions in which a complementary double strand is dissociated.
[claim3]
3. The method according to claim 1, wherein the photo-ligating nucleic acid has a labeling site.
[claim4]
4. The method according to claim 1, wherein the photo-ligating nucleic acid is immobilized onto a support.
[claim5]
5. The method according to claim 1, wherein the light irradiation of the step of inducing photo-ligation is carried out by irradiation with light having a wavelength in the range of 350 to 380 nm.
[claim6]
6. The method according to claim 1, wherein the light irradiation of the step of inducing photo-cleavage is carried out by irradiation with light having a wavelength in the range of 310 to 345 nm.
[claim7]
7. The method according to claim 1, wherein the step of inducing photo-ligation is carried out in a reaction solution containing a salt having a buffering action.
[claim8]
8. A photo-ligating nucleic acid (wherein the nucleic acid includes a nucleic acid and a peptide nucleic acid) having a labeling site and having a group represented by the following formula (I) as a base moiety:
wherein in the formula (I), Ra represents a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen; and
R1 and R2 each independently represent a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen.
[claim9]
9. A photo-ligating nucleic acid (wherein the nucleic acid includes a nucleic acid and a peptide nucleic acid) immobilized onto a support and having a group represented by the following formula (I) as a base moiety:
wherein in the formula (I), Ra represents a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen; and
R1 and R2 each independently represent a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen.
  • 発明者/出願人(英語)
  • FUJIMOTO KENZO
  • YOSHIMURA YOSHINAGA
  • JAPAN ADVANCED INSTITUTE OF SCIENCE AND TECHNOLOGY
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
国際特許分類(IPC)
米国特許分類/主・副
  • C07H021/04
  • C12N015/10A
  • C12Q001/68B10
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