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Title: Optical selection and collection of DNA fragments

Patent ·
OSTI ID:871325
 [1];  [1];  [1];  [1]
  1. Los Alamos, NM
+ Show Author Affiliations

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.

Research Organization:
Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
Assignee:
Regents of University of California (Los Alamos, NM)
Patent Number(s):
US 5707808
OSTI ID:
871325
Country of Publication:
United States
Language:
English

References (10)

The role of dna Damage in pm2 Viral Inactivation by Methylene blue Photosensitization journal May 1994
High-speed photodamage cell selection using bromodeoxyuridine/Hoechst 33342 photosensitized cell killing journal March 1988
Characterization of DNA Size Determination of Small Fragments by Flow Cytometry journal May 1995
Development of a Plaque Reduction Assay and Application to the Study of Psoralen-Damaged dna journal April 1986
Direct observation of tube-like motion of a single polymer chain journal May 1994
Rapid sizing of individual fluorescently stained DNA fragments by flow cytometry journal January 1993
High-speed photodamage cell selection using a frequency-doubled argon ion laser journal March 1995
TOTO and YOYO: New very bright fluorochromes for DNA content analyses by flow cytometry journal February 1994
Stable fluorescent complexes of double-stranded DNA with bis-intercalating asymmetric cyanine dyes: properties and applications journal January 1992
Stable dye–DNA intercalation complexes as reagents for high-sensitivity fluorescence detection journal October 1992

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Related Subjects

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
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