Nanochannel arrays and their preparation and use for high throughput macromolecular analysis
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/68
C12M-001/36
C07H-021/04
출원번호
UP-0484293
(2002-07-25)
등록번호
US-7670770
(2010-04-21)
국제출원번호
PCT/US2002/023610
(2002-07-25)
§371/§102 date
20040120
(20040120)
국제공개번호
WO03/010289
(2003-02-06)
발명자
/ 주소
Chou, Stephen Y.
Cao, Han
Austin, Robert H.
Yu, Zhaoning
Tegenfeldt, Jonas O.
출원인 / 주소
The Trustees of Princeton University
대리인 / 주소
Woodcock Washburn LLP
인용정보
피인용 횟수 :
49인용 특허 :
29
초록▼
Nanochannel arrays that enable high-throughput macromolecular analysis are disclosed. Also disclosed are methods of preparing nanochannel arrays and nanofluidic chips. Methods of analyzing macromolecules, such as entire strands of genomic DNA, are also disclosed, as well as systems for carrying out
Nanochannel arrays that enable high-throughput macromolecular analysis are disclosed. Also disclosed are methods of preparing nanochannel arrays and nanofluidic chips. Methods of analyzing macromolecules, such as entire strands of genomic DNA, are also disclosed, as well as systems for carrying out these methods.
대표청구항▼
We claim: 1. A method of isolating, imaging, and analyzing, in parallel, two or more individual nucleic acid biopolymers, comprising the steps of: providing a nanofluidic chip, comprising: a) nanochannel array, comprising: a surface having a plurality of parallel nanochannels running along the mate
We claim: 1. A method of isolating, imaging, and analyzing, in parallel, two or more individual nucleic acid biopolymers, comprising the steps of: providing a nanofluidic chip, comprising: a) nanochannel array, comprising: a surface having a plurality of parallel nanochannels running along the material of the surface, said nanochannels having a trench width of less than about 150 nanometers and a trench depth of less than 200 nanometers; at least one of the plurality of nanochannels being surmounted by sealing material to render such nanochannels at least substantially enclosed; at least two of the nanochannels capable of admitting a fluid; b) at least one sample reservoir in fluid communication with at least two of the nanochannels, said sample reservoir capable of releasing a fluid containing at least two nucleic acid biopolymer; providing the at least one sample reservoir with at least one fluid, said fluid comprising at least two nucleic acid biopolymers; transporting the at least two nucleic acid biopolymers into the at least two nanochannels to elongate said at least two nucleic acid biopolymers, the at least two nucleic acid biopolymers being individually confined within the at least two nanochannels such that the at least two nucleic acid biopolymer are transported through the at least two nanochannels in an unfolded fashion; imaging in parallel at least one signal transmitted from the at least two elongated and isolated nucleic acid biopolymers within the nanochannels; and correlating the signal to at least one property of the at least two nucleic acid biopolymers to thereby analyze the two or more individual nucleic acid biopolymers. 2. The method according to claim 1 wherein the signal is correlated to at least one of the following properties: length, conformation, and chemical composition. 3. The method of claim 1, wherein the two or more nucleic acid biopolymers are DNA and the signal is correlated to the base pair sequence of said DNA. 4. The method of claim 1 wherein a plurality of reservoirs provide a plurality of nucleic acid biopolymers into a plurality of nanochannels for determining the lengths of the macromolecules. 5. The method of claim 4 wherein more than two of the two or more nucleic acid biopolymers enter a single nanochannel. 6. The method of claim 1, wherein the two or more nucleic acid biopolymers are DNA or RNA. 7. The method of claim 1, wherein the two or more nucleic acid biopolymers are at least substantially unfolded in the channels. 8. The method of claim 1 wherein the concentration of the two or more nucleic acid biopolymers in the fluid is at least one attogram per milliliter. 9. The method of claim 1 wherein the concentration of the two or more nucleic acid biopolymers in the fluid is at least one femtogram per milliliter. 10. The method of claim 1 wherein the concentration of the two or more nucleic acid biopolymers in the fluid is at least one picogram per milliliter. 11. The method of claim 1 wherein the concentration of the two or more nucleic acid biopolymers in the fluid is less than 5 micrograms per milliliter. 12. The method of claim 1 wherein the concentration of the two or more nucleic acid biopolymers in the fluid is less than 0.5 micrograms per milliliter. 13. The method of claim 1 wherein the at least two nucleic acid biopolymers have an elongated length in the channels of greater than 150 nanometers. 14. The method of claim 1 wherein the at least two nucleic acid biopolymers have an elongated length in the channels of greater than 500 nanometers. 15. The method of claim 1 wherein the at least two nucleic acid biopolymers have an elongated length in the channels of greater than 1 micron. 16. The method of claim 1 wherein the at least two nucleic acid biopolymers have an elongated length in the channels of greater than 10 microns. 17. The method of claim 1 wherein the at least two nucleic acid biopolymers are DNA having greater than 10 base pairs. 18. The method of claim 1 wherein the at least two nucleic acid biopolymers are DNA having greater than 100 base pairs. 19. The method of claim 1 wherein the at least two nucleic acid biopolymers are DNA having greater than 1,000 base pairs. 20. The method of claim 1 wherein the at least two nucleic acid biopolymers are DNA having greater than 10,000 base pairs. 21. The method of claim 1 wherein the at least two nucleic acid biopolymers are DNA having greater than 20,000 base pairs. 22. The method of claim 1 wherein the at least two nucleic acid biopolymers are DNA having greater than 40,000 base pairs. 23. The method of claim 1 wherein the at least two nucleic acid biopolymers are DNA having greater than 80,000 base pairs. 24. The method of claim 1 wherein the nanochannel array has at least 96 sample reservoirs for simultaneously analyzing at least 96 different nucleic acid biopolymer fluid samples. 25. The method of claim 1 wherein at least one of the at least two nucleic acid biopolymers are restriction fragment length polymorphisms. 26. The method of claim 1 wherein least one of the at least two nucleic acid biopolymers is a chromosome. 27. The method of claim 26 wherein at least one chromosome is analyzed to determine the presence of at least one single nucleotide polymorphism. 28. The method of claim 1, further comprising the step of illuminating the at least two nucleic acid biopolymers within the at least two nanochannels. 29. The method of claim 1, wherein at least one of the at least two nanochannels is completely enclosed with sealing material on its surface. 30. The method of claim 1, wherein the nanochannel array comprises from more than 2 to about 500,000 parallel channels. 31. The method of claim 1, wherein the sealing material is optically transparent. 32. The method of claim 31, wherein at least one signal is transmitted through the optically transparent sealing material. 33. The method of claim 1, wherein one or more of the plurality of nanochannels is defined by walls that span the length of the substrate surface.
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