Raman-based system for DNA sequencing-mapping and other separations

• US 5,306,403 A
• Filed: 08/24/1992
• Issued: 04/26/1994
• Est. Priority Date: 08/24/1992
• Status: Expired due to Fees

• Current Assignee: Martin Marietta Energy Systems, Inc.
• Original Assignee: Martin Marietta Energy Systems, Inc.
• Inventors: Vo-Dinh, Tuan
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US07/933,376
• Time in Patent Office: 610 Days
• Field of Search: 204/299 R, 204/182.8, 204/180.1, 73/61.58, 73/51.54
• US Class Current: 204/450
• CPC Class Codes: G01N 21/658 enhancement Raman, e.g. surface plasmons
• CPC Class Codes: G01N 27/44721 by optical means

Abstract

DNA sequencing and mapping are performed by using a Raman spectrometer with a surface enhanced Raman scattering (SERS) substrate to enhance the Raman signal. A SERS label is attached to a DNA fragment and then analyzed with the Raman spectrometer to identify the DNA fragment according to characteristics of the Raman spectrum generated.

Claims

1. An apparatus for analyzing DNA comprising:

a SERS label attached to DNA fragments;

sequencer means for separating the labeled DNA fragments;

a SERS-active media, disposed within the sequencer means, and having at least one of the DNA fragments adsorbed thereon; and

Raman spectrometer means, including a light source focused onto the SERS-active media, for producing a SERS spectrum of the SERS labeled DNA fragment, the SERS spectrum having characteristics which identify the DNA fragment.

2. An apparatus according to claim 1, wherein the sequencer means a gel electrophoresis apparatus having an enclosure, the enclosure including two parallel glass plates, and the SERS-active media is a coating disposed on an inner surface of at least one of the two glass plates.

3. An apparatus according to claim 2, wherein the SERS-active coating is silver-coated TiO₂.

4. An apparatus according to claim 1, wherein the sequencer means a gel electrophoresis apparatus having an enclosure containing a gel, the enclosure including two parallel glass plates, and the SERS-active media are metal microparticles disposed in the gel.

5. An apparatus according to claim 1, wherein the SERS label is a dye selected from the group consisting of aminoacridine, TRIT, NBD, fluorescein, and Texas Red dye.

6. An apparatus according to claim 1, wherein the SERS label is selected from a chemical group capable of specifically attaching to one of a sugar group, a base and a phosphate group of a DNA nucleotide.

7. An apparatus for analyzing DNA comprising:

a SERS label attached to DNA fragments;

sequencing means for separating the SERS labeled DNA fragments;

means for transferring the SERS labeled DNA fragments from the sequencer means onto a moving SERS-active substrate; and

Raman spectrometer means, having a light source focused onto the moving SERS-active substrate, for producing a SERS spectrum of the SERS labeled DNA fragment, the SERS spectrum having characteristics which identify the DNA fragment.

8. An apparatus according to claim 7, wherein the transferring means is an automated blotting apparatus.

9. A method for analyzing DNA comprising:

attaching a SERS label to DNA fragments;

separating the labeled DNA fragments with a sequencer;

adsorbing at least one of the DNA fragments onto a SERS-active media disposed within the sequencer; and

producing a SERS spectrum of the SERS labeled DNA fragment with a Raman spectrometer, the SERS spectrum having characteristics which identify the DNA fragment.

10. A method according to claim 9, wherein separating the labeled DNA fragments comprises passing the DNA fragments through a gel electrophoresis apparatus.

11. A method according to claim 9, wherein separating the labeled DNA fragments comprises passing the DNA fragments through a capillary electrophoresis apparatus.

12. A method according to claim 9, wherein separating the labeled DNA fragments comprises passing the DNA fragments through a thin-layer electrophoresis apparatus.

13. A method according to claim 9, wherein separating the labeled DNA fragments comprises passing the DNA fragments through a paper electrophoresis apparatus.

14. A method according to claim 9, wherein separating the labeled DNA fragments comprises passing the DNA fragments through a thin-layer chromatography apparatus.

15. A method according to claim 9, wherein separating the labeled DNA fragments comprises passing the DNA fragments through a paper chromatography apparatus.