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Title: Automated hybridization/imaging device for fluorescent multiplex DNA sequencing

Abstract

A method is disclosed for automated multiplex sequencing of DNA with an integrated automated imaging hybridization chamber system. This system comprises an hybridization chamber device for mounting a membrane containing size-fractionated multiplex sequencing reaction products, apparatus for fluid delivery to the chamber device, imaging apparatus for light delivery to the membrane and image recording of fluorescence emanating from the membrane while in the chamber device, and programmable controller apparatus for controlling operation of the system. The multiplex reaction products are hybridized with a probe, the enzyme (such as alkaline phosphatase) is bound to a binding moiety on the probe, and a fluorogenic substrate (such as a benzothiazole derivative) is introduced into the chamber device by the fluid delivery apparatus. The enzyme converts the fluorogenic substrate into a fluorescent product which, when illuminated in the chamber device with a beam of light from the imaging apparatus, excites fluorescence of the fluorescent product to produce a pattern of hybridization. The pattern of hybridization is imaged by a CCD camera component of the imaging apparatus to obtain a series of digital signals. These signals are converted by the controller apparatus into a string of nucleotides corresponding to the nucleotide sequence an automated sequencemore » reader. The method and apparatus are also applicable to other membrane-based applications such as colony and plaque hybridization and Southern, Northern, and Western blots. 9 figs.« less

Inventors:
; ; ; ; ; ;
Issue Date:
Research Org.:
Univ. of Utah, Salt Lake City, UT (United States)
OSTI Identifier:
131929
Patent Number(s):
5,470,710
Application Number:
PAN: 8-141,234; CNN: Grant R01-HG00517-02;Grant P30-HG00199-03
Assignee:
Univ. of Utah, Salt Lake City, UT (United States)
DOE Contract Number:  
FG02-88ER60700
Resource Type:
Patent
Resource Relation:
Other Information: PBD: 28 Nov 1995
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; DNA SEQUENCING; IMAGE PROCESSING; CAMERAS; DESIGN; DNA HYBRIDIZATION; BIOCHEMICAL REACTION KINETICS; ENZYMES

Citation Formats

Weiss, R B, Kimball, A W, Gesteland, R F, Ferguson, F M, Dunn, D M, Di Sera, L J, and Cherry, J L. Automated hybridization/imaging device for fluorescent multiplex DNA sequencing. United States: N. p., 1995. Web.
Weiss, R B, Kimball, A W, Gesteland, R F, Ferguson, F M, Dunn, D M, Di Sera, L J, & Cherry, J L. Automated hybridization/imaging device for fluorescent multiplex DNA sequencing. United States.
Weiss, R B, Kimball, A W, Gesteland, R F, Ferguson, F M, Dunn, D M, Di Sera, L J, and Cherry, J L. Tue . "Automated hybridization/imaging device for fluorescent multiplex DNA sequencing". United States.
@article{osti_131929,
title = {Automated hybridization/imaging device for fluorescent multiplex DNA sequencing},
author = {Weiss, R B and Kimball, A W and Gesteland, R F and Ferguson, F M and Dunn, D M and Di Sera, L J and Cherry, J L},
abstractNote = {A method is disclosed for automated multiplex sequencing of DNA with an integrated automated imaging hybridization chamber system. This system comprises an hybridization chamber device for mounting a membrane containing size-fractionated multiplex sequencing reaction products, apparatus for fluid delivery to the chamber device, imaging apparatus for light delivery to the membrane and image recording of fluorescence emanating from the membrane while in the chamber device, and programmable controller apparatus for controlling operation of the system. The multiplex reaction products are hybridized with a probe, the enzyme (such as alkaline phosphatase) is bound to a binding moiety on the probe, and a fluorogenic substrate (such as a benzothiazole derivative) is introduced into the chamber device by the fluid delivery apparatus. The enzyme converts the fluorogenic substrate into a fluorescent product which, when illuminated in the chamber device with a beam of light from the imaging apparatus, excites fluorescence of the fluorescent product to produce a pattern of hybridization. The pattern of hybridization is imaged by a CCD camera component of the imaging apparatus to obtain a series of digital signals. These signals are converted by the controller apparatus into a string of nucleotides corresponding to the nucleotide sequence an automated sequence reader. The method and apparatus are also applicable to other membrane-based applications such as colony and plaque hybridization and Southern, Northern, and Western blots. 9 figs.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1995},
month = {11}
}