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:

Roslaniec, Mary C ^[1]; Martin, John C ^[1]; Jett, James H ^[1]; Cram, L Scott ^[1]

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+ Show Inventor Affiliations

1. Los Alamos, NM

Issue Date:

1998-01-13

Research Org.:

Los Alamos National Laboratory (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):

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

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C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N15/1006 - {by means of a solid support carrier, e.g. particles, polymers}

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

C12Q1/68 - involving nucleic acids

G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES

G01N15/1459 - {the analysis being performed on a sample stream}
G01N2015/0038 - {Investigating nanoparticles}
G01N2015/149 - {Sorting the particles}

Y - NEW / CROSS SECTIONAL TECHNOLOGIES Y10 - TECHNICAL SUBJECTS COVERED BY FORMER USPC Y10T - TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

Y10T436/143333 - Saccharide [e.g., DNA, etc.]

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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/