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Sequence-specific nucleic acid purification method manner

Foreign code F110005606
File No. RX02P38WO
Posted date Sep 9, 2011
Country United States of America
Application number 93747109
Gazette No. 20110034683
Gazette No. 7972792
Date of filing Apr 16, 2009
Gazette Date Feb 10, 2011
Gazette Date Jul 5, 2011
International application number JP2009001747
International publication number WO2009128266
Date of international filing Apr 16, 2009
Date of international publication Oct 22, 2009
Priority data
  • P2008-106642 (Apr 16, 2008) JP
  • 2009WO-JP01747 (Apr 16, 2009) WO
Title Sequence-specific nucleic acid purification method manner
Abstract (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.
Scope of claims [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.
  • Inventor, and Inventor/Applicant
  • FUJIMOTO KENZO
  • YOSHIMURA YOSHINAGA
  • JAPAN ADVANCED INSTITUTE OF SCIENCE AND TECHNOLOGY
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
U.S. Cl./(Sub)
  • C07H021/04
  • C12N015/10A
  • C12Q001/68B10
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