US5230781A — Sequencing near infrared and infrared fluorescenc…

Claim 1

Independent Claim 176 words

A method for sequencing DNA comprising: applying DNA samples marked with fluorescent dye having absorbance and fluorescense maxima at near infrared or infrared wavelengths when combined with the DNA at a plurality of locations in a gel electrophoresis slab for electrophoresing in a plurality of channels through a gel electrophoresis slab; establishing electrical potential across said gel electrophoresis slab wherein DNA samples are resolved in accordance with the size of DNA fragments in said gel electrophoresis slab into fluorescently marked DNA bands; scanning the separated samples photoelectrically while they are in the slab with a laser diode and a sensor wherein the laser scans with near infrared or infrared scanning light at a scanning light frequency within the absorbance spectrum of said fluorescently marked DNA samples and sensing light at the emission frequency of the marked DNA; modulating said light from said laser at a predetermined modulation frequency; detecting fluorescent light emitted by said DNA bands at said modulation frequency, whereby background noise from the medium through which the light is transmitted may be discriminated against.

Claim 2

Independent Claim 176 words

A method for sequencing DNA comprising: applying DNA samples marked with fluorescent dye having absorbance and fluorescense maxima at near infrared or infrared wavelengths when combined with the DNA at a plurality of locations in a gel electrophoresis slab for electrophoresing in a plurality of channels through a gel electrophoresis slab; establishing electrical potential across said gel electrophoresis slab wherein DNA samples are resolved in accordance with the size of DNA fragments in said gel electrophoresis slab into fluorescently marked DNA bands; scanning the separated samples photoelectrically while they are in the slab with a laser diode and a sensor wherein the laser scans with near infrared or infrared scanning light at a scanning light frequency within the absorbance spectrum of said fluorescently marked DNA samples and sensing light at the emission frequency of the marked DNA; modulating said light from said laser at a predetermined modulation frequency; detecting fluorescent light emitted by said DNA bands at said modulation frequency, whereby background noise from the medium through which the light is transmitted may be discriminated against.

Claim 3

Dependent Claim 24 words

A method according to claim 1 in which the light is modulated at a frequency within the band between 100 hz to 30 KH.

Claim 4

Dependent Claim 76 words

A method according to claim 2 further including means for resolving the bands within at least one channel so that the bands of more mobile strands in the channel are fully resolved while some of the less mobile strands to be later formed into bands are unresolved in a continuous process such that at least 10 percent of the bands are fully resolved while the less mobile strands are yet unresolved into bands in the channel.

Claim 5

Independent Claim 147 words

A method of DNA sequencing comprising the steps of: applying opposite polarity electrical potentials to a first and at least second buffer; applying fluorescently marked DNA strands to a plurality of channels of gel, whereby said fluorescently marked DNA is electrophoresed along said gel so that the bands of the more mobile strands in at least one channel is fully resolved while some of the less mobile strands to be later formed into bands are unresolved in a continuous process; scanning across said channels with light emitted from a laser; and detecting fluorescent light emitted by said fluorescently marked strands, whereby the time sequence of separated bands may be obtained, wherein the light from said laser is in the band incorporating at least the near infrared and infrared regions and said detector responds to light in a band including at least said near infrared and infrared regions.

Claim 6

Independent Claim 147 words

A method of DNA sequencing comprising the steps of: applying opposite polarity electrical potentials to a first and at least second buffer; applying fluorescently marked DNA strands to a plurality of channels of gel, whereby said fluorescently marked DNA is electrophoresed along said gel so that the bands of the more mobile strands in at least one channel is fully resolved while some of the less mobile strands to be later formed into bands are unresolved in a continuous process; scanning across said channels with light emitted from a laser; and detecting fluorescent light emitted by said fluorescently marked strands, whereby the time sequence of separated bands may be obtained, wherein the light from said laser is in the band incorporating at least the near infrared and infrared regions and said detector responds to light in a band including at least said near infrared and infrared regions.

Claim 7

Dependent Claim 143 words

Apparatus for sequencing DNA comprising: a gel electrophoresis slab; means for establishing electrical potential across gel electrophoresis slab; means for applying DNA samples at a plurality of locations for electrophoresing in a plurality of channels through said gel electrophoresis slab; means for separating DNA samples in accordance with the size of DNA fragments in said gel electrophoresis slab; means for scanning the separated samples photoelectrically; said means for scanning including a laser diode having an emission spectrum that includes one or more wavelengths in the infrared region and near infrared region and a sensor; a modulator; a detector being positioned to absorb fluorescent light emitted by said DNA bands; said sensor being applied to means for detecting pulses of light having the same pulse rate as laser light pulses, whereby background noise from the medium through which it is applied is discriminated against.

Claim 8

Dependent Claim 143 words

Apparatus for sequencing DNA comprising: a gel electrophoresis slab; means for establishing electrical potential across gel electrophoresis slab; means for applying DNA samples at a plurality of locations for electrophoresing in a plurality of channels through said gel electrophoresis slab; means for separating DNA samples in accordance with the size of DNA fragments in said gel electrophoresis slab; means for scanning the separated samples photoelectrically; said means for scanning including a laser diode having an emission spectrum that includes one or more wavelengths in the infrared region and near infrared region and a sensor; a modulator; a detector being positioned to absorb fluorescent light emitted by said DNA bands; said sensor being applied to means for detecting pulses of light having the same pulse rate as laser light pulses, whereby background noise from the medium through which it is applied is discriminated against.

Claim 9

Dependent Claim 56 words

Apparatus for sequencing DNA according to claim 5 in which said modulator is positioned between said laser and said DNA bands, whereby said light from said laser is pulsed at a predetermined rate, and said scanning apparatus is aligned with said gel, whereby the bands are scanned while still in the gel but after being resolved.

Claim 10

Dependent Claim 182 words

DNA sequencing apparatus comprising gel electrophoresis means; one end of said gel electrophoresis means communicating with a buffer solution; at least one other side of said gel electrophoresis means communicating with a second buffer solution; means for applying opposite polarity electrical potentials to said first and at least second buffer; means for applying fluorescently marked DNA strands to a plurality of channels of said gel, whereby said fluorescently marked DNA is electrophoresed along said gel so that the bands of the more mobile strands in at least one channel is fully resolved while some of the less mobile strands to be later formed into bands are unresolved in a continuous process; means for scanning across said channels with light emitted from a laser diode, wherein said light from said laser in in the band incorporates at least the near infrared and infrared regions; said detector responds to light in a band including at least said near infrared and infrared regions; and means for detecting fluorescent light emitted by said fluorescently marked strands, whereby the time sequence of separated bands may be obtained.

Claim 11

Dependent Claim 182 words

DNA sequencing apparatus comprising gel electrophoresis means; one end of said gel electrophoresis means communicating with a buffer solution; at least one other side of said gel electrophoresis means communicating with a second buffer solution; means for applying opposite polarity electrical potentials to said first and at least second buffer; means for applying fluorescently marked DNA strands to a plurality of channels of said gel, whereby said fluorescently marked DNA is electrophoresed along said gel so that the bands of the more mobile strands in at least one channel is fully resolved while some of the less mobile strands to be later formed into bands are unresolved in a continuous process; means for scanning across said channels with light emitted from a laser diode, wherein said light from said laser in in the band incorporates at least the near infrared and infrared regions; said detector responds to light in a band including at least said near infrared and infrared regions; and means for detecting fluorescent light emitted by said fluorescently marked strands, whereby the time sequence of separated bands may be obtained.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1996-07-26	1997-01-25	1997-07-27	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	2000-07-26	2001-01-25	2001-07-27	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2004-07-26	2005-01-25	2005-07-27	$7700.00	Window Closed / Lapsed

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

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