Sequencing near infrared and infrared fluorescence labeled DNA for detecting using laser diodes and suitable labels therefor

• US 6,086,737 A
• Filed: 07/11/1995
• Issued: 07/11/2000
• Est. Priority Date: 03/29/1984
• Status: Expired due to Fees

• Current Assignee: Li & S Company Limited (Yesco Holdings Co., Ltd.)
• Original Assignee: Li & S Company Limited (Yesco Holdings Co., Ltd.)
• Inventors: Patonay, Gabor, Narayanan, Narasimhachari, Middendorf, Lyle Richard, Brumbaugh, John A.
• Primary Examiner(s): Beisner, William H.
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US08/500,691
• Time in Patent Office: 1,827 Days
• Field of Search: 204/299 R, 204/182.8, 204/180.1, 204/461, 204/466, 204/467, 204/456, 204/452, 204/451, 204/612, 204/616, 204/618, 204/606, 204/603, 204/601, 436/800, 536/25.32, 435/6, 435/968
• US Class Current: 204/461
• CPC Class Codes: C07D 209/12 Radicals substituted by oxygen atoms
• CPC Class Codes: C07D 209/62 Naphtho [c] pyrroles; Hydrogenated naphtho [c] pyrroles
• CPC Class Codes: C09B 23/0025 the substituent being bound through an oxygen atom
• CPC Class Codes: C09B 23/0066 the polymethine chain being part of a carbocyclic ring,(e.g. benzene, naphtalene, cyclohexene, cyclobutenene-quadratic acid)
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 27/447 using electrophoresis
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44726 using specific dyes, markers or binding molecules
• CPC Class Codes: Y10S 435/968 High energy substrates, e.g. fluorescent, chemiluminescent, radioactive
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

To sequence DNA automatically, DNA marked with far infrared, near infrared, or infrared fluorescent dyes are electrophoresed in a plurality of channels through a gel electrophoresis slab or capillary tubes wherein the DNA samples are resolved in accordance with the size of DNA fragments in the gel electrophoresis slab or capillary tubes into fluorescently marked DNA bands. The separated samples are scanned photoelectrically with a laser diode and a sensor, wherein the laser scans with scanning light at a wavelength within the absorbance spectrum of said fluorescently marked DNA samples and light is sensed at the emission wavelength of the marked DNA.

Claims

1. A method of identifying strands of DNA comprising the steps of:

marking the strands with fluorescent labels that emit light in a region of wavelengths including at least one wavelength within one of a far red, near infrared, and infrared regions wherein the fluorescent label includes a chromophore having the formula;

##STR7## Where X is H or OCH₃ ;

irradiating the strands with light having the wavelength within one of the far red, near infrared, and infrared regions; and

detecting the light emitted from the fluorescent labels.

2. A method of identifying strands of DNA in accordance with claim 1 in which the step of detecting includes the step of detecting one of far red, near infrared, and infrared light emissions from the fluorescent labels.

3. A method of identifying strands of DNA comprising the steps of:

marking the strands with fluorescent labels that emit light in a region of wavelengths including at least one wavelength within one of a far red, near infrared, and infrared regions wherein the fluorescent label includes a chromophore having the formula;

##STR8## where R is ethyl or 4-sulfonatobutyl and n is either 1 or 2;

irradiating the strands with light having a wavelength within one of the far red, near infrared, and infrared regions; and

detecting the light emitted from the fluorescent labels.

4. A method of identifying strands of DNA in accordance with claim 3 in which the step of detecting includes the step of detecting one of far red, near infrared, and infrared light emissions from the fluorescent labels.

5. A method of identifying strands of DNA comprising the steps of:

marking the strands with fluorescent labels that emit light in a region of wavelengths including at least one wavelength within one of a far red, near infrared, and infrared regions wherein the fluorescent label includes a chromophore having the formula;

##STR9## irradiating the strands with light having the wavelength within one of the far red, near infrared, and infrared regions; and

detecting the light emitted from the fluorescent labels.

6. A method of identifying strands of DNA in accordance with claim 5 in which the step of detecting includes the step of detecting one of far red, near infrared and infrared light emissions from the fluorescent labels.

7. A method of identifying strands of DNA comprising the steps of:

marking the strands with fluorescent labels that emit light in a region of wavelengths including at least one wavelength within one of a far red, near infrared, and infrared region wherein the fluorescent label includes a chromophore having the formula;

##STR10## irradiating the strands with light having the wavelength within one of the far red, near infrared, and infrared regions; and

detecting the light emitted from the fluorescent labels.

8. A method of identifying strands of DNA in accordance with claim 7 in which the step of detecting includes the step of detecting one of far red, near infrared, and infrared light emissions from the fluorescent labels.

9. A method of identifying strands of DNA comprising the steps of:

marking the strands with fluorescent labels that emit light in a region of wavelengths including at least one wavelength within one of a far red, near infrared, and infrared regions wherein the fluorescent label includes a chromophore having the formula;

##STR11## applying the DNA strands marked with fluorescent labels at a plurality of locations in a gel electrophoresis slab or set of capillaries for electrophoresing in a plurality of channels through the gel electrophoresis slab or set of capillaries wherein the fluorescent labels include a fluorescent dye having absorbance and fluorescense maxima at one of the far red, near infrared and infrared wavelengths when combined with DNA;

establishing electrical potential across said gel electrophoresis slab or set of capillaries wherein the marked DNA strands are resolved into bands of fluorescently marked DNA strands in said gel electrophoresis slab or set of capillaries in accordance with the size of the fluorescently marked DNA strands; and

irradiating the bands of fluorescently marked DNA strands with light having the wavelength within one of the far red, near infrared, and infrared regions while they are in the slab or set of capillaries with a laser diode wherein the laser diode scans with far red, near infrared, or infrared scanning light at a wavelength within an absorbance spectrum of said fluorescently marked DNA strands; and

detecting the light emitted from the fluorescent labels with a sensor wherein the sensor senses light at the emission wavelength of the marked DNA.