Multiplex analysis of DNA
Abstract
This invention features vectors and a method for sequencing DNA. The method includes the steps of: a) ligating the DNA into a vector comprising a tag sequence, the tag sequence includes at least 15 bases, wherein the tag sequence will not hybridize to the DNA under stringent hybridization conditions and is unique in the vector, to form a hybrid vector, b) treating the hybrid vector in a plurality of vessels to produce fragments comprising the tag sequence, wherein the fragments differ in length and terminate at a fixed known base or bases, wherein the fixed known base or bases differs in each vessel, c) separating the fragments from each vessel according to their size, d) hybridizing the fragments with an oligonucleotide able to hybridize specifically with the tag sequence, and e) detecting the pattern of hybridization of the tag sequence, wherein the pattern reflects the nucleotide sequence of the DNA.
- Inventors:
-
- Boston, MA
- Dorchester, MA
- Issue Date:
- Research Org.:
- Harvard University
- OSTI Identifier:
- 868474
- Patent Number(s):
- 5149625
- Assignee:
- President and Fellows of Harvard College (Cambridge, MA)
- Patent Classifications (CPCs):
-
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
Y - NEW / CROSS SECTIONAL TECHNOLOGIES Y10 - TECHNICAL SUBJECTS COVERED BY FORMER USPC Y10S - TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- DOE Contract Number:
- FG02-87ER60565
- Resource Type:
- Patent
- Country of Publication:
- United States
- Language:
- English
- Subject:
- multiplex; analysis; dna; features; vectors; method; sequencing; steps; ligating; vector; comprising; tag; sequence; 15; bases; hybridize; stringent; hybridization; conditions; unique; form; hybrid; treating; plurality; vessels; produce; fragments; length; terminate; fixed; base; differs; vessel; separating; according; size; hybridizing; oligonucleotide; specifically; detecting; pattern; reflects; nucleotide; tag sequence; sequencing dna; nucleotide sequence; vector comprising; /435/436/
Citation Formats
Church, George M, and Kieffer-Higgins, Stephen. Multiplex analysis of DNA. United States: N. p., 1992.
Web.
Church, George M, & Kieffer-Higgins, Stephen. Multiplex analysis of DNA. United States.
Church, George M, and Kieffer-Higgins, Stephen. Wed .
"Multiplex analysis of DNA". United States. https://www.osti.gov/servlets/purl/868474.
@article{osti_868474,
title = {Multiplex analysis of DNA},
author = {Church, George M and Kieffer-Higgins, Stephen},
abstractNote = {This invention features vectors and a method for sequencing DNA. The method includes the steps of: a) ligating the DNA into a vector comprising a tag sequence, the tag sequence includes at least 15 bases, wherein the tag sequence will not hybridize to the DNA under stringent hybridization conditions and is unique in the vector, to form a hybrid vector, b) treating the hybrid vector in a plurality of vessels to produce fragments comprising the tag sequence, wherein the fragments differ in length and terminate at a fixed known base or bases, wherein the fixed known base or bases differs in each vessel, c) separating the fragments from each vessel according to their size, d) hybridizing the fragments with an oligonucleotide able to hybridize specifically with the tag sequence, and e) detecting the pattern of hybridization of the tag sequence, wherein the pattern reflects the nucleotide sequence of the DNA.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Wed Jan 01 00:00:00 EST 1992},
month = {Wed Jan 01 00:00:00 EST 1992}
}