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Title: Optical selection and collection of DNA fragments

Abstract

Optical selection and collection of DNA fragments. The present invention includes the optical selection and collection of large (>.mu.g) quantities of clonable, chromosome-specific DNA from a sample of chromosomes. Chromosome selection is based on selective, irreversible photoinactivation of unwanted chromosomal DNA. Although more general procedures may be envisioned, the invention is demonstrated by processing chromosomes in a conventional flow cytometry apparatus, but where no droplets are generated. All chromosomes in the sample are first stained with at least one fluorescent analytic dye and bonded to a photochemically active species which can render chromosomal DNA unclonable if activated. After passing through analyzing light beam(s), unwanted chromosomes are irradiated using light which is absorbed by the photochemically active species, thereby causing photoinactivation. As desired chromosomes pass this photoinactivation point, the inactivating light source is deflected by an optical modulator; hence, desired chromosomes are not photoinactivated and remain clonable. The selection and photoinactivation processes take place on a microsecond timescale. By eliminating droplet formation, chromosome selection rates 50 times greater than those possible with conventional chromosome sorters may be obtained. Thus, usable quantities of clonable DNA from any source thereof may be collected.

Inventors:
 [1];  [1];  [1];  [1]
  1. Los Alamos, NM
Issue Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
OSTI Identifier:
871325
Patent Number(s):
5707808
Assignee:
Regents of University of California (Los Alamos, NM)
Patent Classifications (CPCs):
G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
Y - NEW / CROSS SECTIONAL TECHNOLOGIES Y10 - TECHNICAL SUBJECTS COVERED BY FORMER USPC Y10T - TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
optical; selection; collection; dna; fragments; quantities; clonable; chromosome-specific; sample; chromosomes; chromosome; based; selective; irreversible; photoinactivation; unwanted; chromosomal; procedures; envisioned; demonstrated; processing; conventional; flow; cytometry; apparatus; droplets; generated; stained; fluorescent; analytic; dye; bonded; photochemically; active; species; render; unclonable; activated; passing; analyzing; light; beam; irradiated; absorbed; causing; desired; pass; inactivating; source; deflected; modulator; hence; photoinactivated; remain; processes; microsecond; timescale; eliminating; droplet; formation; rates; 50; times; sorters; obtained; usable; collected; droplet formation; photochemically active; chromosomal dna; light source; light beam; dna fragments; flow cytometry; dna fragment; optical selection; chemically active; optical modulator; active species; activation process; usable quantities; specific dna; cytometry apparatus; /435/436/

Citation Formats

Roslaniec, Mary C, Martin, John C, Jett, James H, and Cram, L Scott. Optical selection and collection of DNA fragments. United States: N. p., 1998. Web.
Roslaniec, Mary C, Martin, John C, Jett, James H, & Cram, L Scott. Optical selection and collection of DNA fragments. United States.
Roslaniec, Mary C, Martin, John C, Jett, James H, and Cram, L Scott. Tue . "Optical selection and collection of DNA fragments". United States. https://www.osti.gov/servlets/purl/871325.
@article{osti_871325,
title = {Optical selection and collection of DNA fragments},
author = {Roslaniec, Mary C and Martin, John C and Jett, James H and Cram, L Scott},
abstractNote = {Optical selection and collection of DNA fragments. The present invention includes the optical selection and collection of large (>.mu.g) quantities of clonable, chromosome-specific DNA from a sample of chromosomes. Chromosome selection is based on selective, irreversible photoinactivation of unwanted chromosomal DNA. Although more general procedures may be envisioned, the invention is demonstrated by processing chromosomes in a conventional flow cytometry apparatus, but where no droplets are generated. All chromosomes in the sample are first stained with at least one fluorescent analytic dye and bonded to a photochemically active species which can render chromosomal DNA unclonable if activated. After passing through analyzing light beam(s), unwanted chromosomes are irradiated using light which is absorbed by the photochemically active species, thereby causing photoinactivation. As desired chromosomes pass this photoinactivation point, the inactivating light source is deflected by an optical modulator; hence, desired chromosomes are not photoinactivated and remain clonable. The selection and photoinactivation processes take place on a microsecond timescale. By eliminating droplet formation, chromosome selection rates 50 times greater than those possible with conventional chromosome sorters may be obtained. Thus, usable quantities of clonable DNA from any source thereof may be collected.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1998},
month = {1}
}

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Works referenced in this record:

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journal, May 1994


High-speed photodamage cell selection using bromodeoxyuridine/Hoechst 33342 photosensitized cell killing
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