EP0630972A2
Expired / Lapsed
EP
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DNA analyzing method
Assignee: Hitachi Ltd
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Filing: 1994-06-23
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Granted: —
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Expires: 2014-06-23
Abstract
Abstract A DNA analyzing method which bonds the oligomer of the known base sequence to the DNA fragment obtained by digesting sample 1 with restrictive enzyme, causes hybridization between oligomer 3 and DNA fragment using the oligomers 3 which have the sequences of all combinations of the types of the bases within the length of several bases following the known base sequence, checks presence or absence of the hybridization or complementary DNA strand extension, identifies the DNA fragment terminal sequence from this result, and fractionates the DNA fragments and analyzes them or analyzes them as they are. This DNA analyzing method provides an effective analysis of mixtures of long DNAs or DNA fragments.
Inventors
- 👤 Hideki Kambara
- 👤 Kazunori Okano
- 👤 Satoshi Takahashi
- 👤 Keiichi Nagai
- 👤 Kazuko Nr. 309 Hitachi Ozaki-Haitsu Kawamoto
- 👤 Hiroko Nr. 110 Hitachi Ozaki-Haitsu Furuyama
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-09 21:28
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6869; C12Q1/6844; C12Q1/6846
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
65 words
A DNA analyzing method comprising: i) a process step of forming DNA fragments by digesting a DNA strand, ii) a process step of recognizing the base sequences in the vicinity of the terminals of DNA fragments, and fractionating or collecting the DNA fragments separately, and iii) a process step of sequencing each of DNA fragments recognized and fractionated or separately collected in process step ii).
Claim 2
Dependent Claim
115 words
A DNA analyzing method comprising I) a process step of digesting the double stranded DNA, II) a process step of bonding a DNA oligomer having a known base sequence to the digested portion of the DNA, III) a process step of recognizing the base sequence of the DNA which follows the known base sequence at the terminal of the DNA after having carried through process step II), and fractionating or collecting the digested DNA fragments separately according to the difference of the base sequence in the vicinity of the terminals of the DNA fragments, and IV) a process step of determining the base sequence of the DNA fractionated or separately collected in process step III.
Claim 3
Dependent Claim
54 words
A DNA analyzing method according to Claim 1, wherein, in process step ii), oligomers having different base sequences are each fixed separately on an array sensor for each base sequence of oligomer, and the base sequences in the vicinity of the terminals of said DNA fragments are recognized with the oligomer fixed array sensor.
Claim 4
Dependent Claim
41 words
A DNA analyzing method according to Claim 3, wherein, said array sensor in the process ii) comprises a combination of a plurality of movable line sensors in which each of oligomers having various sequence is immobilized in a cell arrayed linearly.
Claim 5
Dependent Claim
59 words
A DNA analyzing method according to any of the Claims 1, 3 and 4, which includes, between said process steps i) and ii), another process step i') of bonding the terminal of the DNA fragment with DNAs having known base sequences which are linked with an agent selected from the group consisting of fluorophore, electroluminescence reagent and radioisotope label.
Claim 6
Dependent Claim
52 words
A DNA analyzing method according to Claim 5, which, in said process step ii), allows recognition of the base sequence in the vicinity of the DNA fragment terminal by hybridization between the vicinity of said DNA fragment including, at least, part of the known base sequence and oligomer on the array sensor.
Claim 7
Dependent Claim
86 words
A DNA analyzing method according to Claim 6, wherein, in said process step ii), the base sequence of the oligomer on the array sensor is complementary to the base sequence comprising: a) base sequence of, at least, part of the labeled DNA having the known base sequence b) base sequence of the recognition sequence recognized by the restriction enzyme used to digest the DNA in process i), and c) the base sequence having at least two-base length for identifying DNA fragments, following the enzyme recognizing sequence.
Claim 8
Dependent Claim
62 words
A DNA analyzing method according to Claim 6, which, in said process ii), allows detection of the presence or absence of hybridization between said DNA fragment and oligomer on the array sensor, recognition of the base sequence at the terminal of the DNA fragment according to the result, and fractionation, separate taking or analysis of said DNA fragment on the array sensor.
Claim 9
Dependent Claim
21 words
A DNA analyzing method according to Claim 7, wherein said base sequence c) has a length of 2 to 6 mer.
Claim 10
Dependent Claim
43 words
A DNA analyzing method according to Claim 2, which includes between said process steps II) and III), another process step II') of converting at least the portion in the vicinity of the terminal of said double stranded DNA into a single stranded form.
Claim 11
Dependent Claim
60 words
A DNA analyzing method according to Claim 2 or Claim 10, wherein, in said process step III), more than one type of oligomer having different base sequences is separated for each base sequence, and the base sequence of the DNA which follows the known base sequence at the terminals of said DNA fragments is recognized on a fixed array sensor.
Claim 12
Dependent Claim
62 words
A DNA analyzing method comprising A) a process step of digesting double stranded DNA, B) a process step of bonding an oligomer having a known base sequence to the digested portion of the DNA, and C) a process step of recognizing the base sequence portion of the DNA following the known base sequence, and synthesizing the DNA complementary to the specific fragment.
Claim 13
Dependent Claim
35 words
A DNA analyzing method according to Claim 3 or Claim 11 wherein, in said process step ii) or III), the base sequence in the vicinity of the terminal is recognized through complementary DNA strand extension.
References Cited (Backward Citations)
No citation records indexed.
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