Single molecule arrays for genetic and chemical analysis
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/68
C07H-021/04
C07K-001/04
출원번호
US-0714133
(2015-05-15)
등록번호
US-9650673
(2017-05-16)
발명자
/ 주소
Drmanac, Radoje
Callow, Matthew J.
Drmanac, Snezana
Hauser, Brian K.
Yeung, George
출원인 / 주소
Complete Genomics, Inc.
대리인 / 주소
Kilpatrick Townsend & Stockton LLP
인용정보
피인용 횟수 :
1인용 특허 :
110
초록▼
Random arrays of single molecules are provided for carrying out large scale analyzes, particularly of biomolecules, such as genomic DNA, cDNAs, proteins, and the like. In one aspect, arrays of the invention comprise concatemers of DNA fragments that are randomly disposed on a regular array of discre
Random arrays of single molecules are provided for carrying out large scale analyzes, particularly of biomolecules, such as genomic DNA, cDNAs, proteins, and the like. In one aspect, arrays of the invention comprise concatemers of DNA fragments that are randomly disposed on a regular array of discrete spaced apart regions, such that substantially all such regions contain no more than a single concatemer. Preferably, such regions have areas substantially less than 1 μm2 and have nearest neighbor distances that permit optical resolution of on the order of 109 single molecules per cm2. Many analytical chemistries can be applied to random arrays of the invention, including sequencing by hybridization chemistries, sequencing by synthesis chemistries, SNP detection chemistries, and the like, to greatly expand the scale and potential applications of such techniques.
대표청구항▼
1. An array comprising: (a) a solid substrate comprising a surface, wherein said surface comprises a plurality of discrete regions defined on said surface,wherein said discrete regions are arranged on the surface in a regular array and are separated by inter-regional areas;(b) a population of concat
1. An array comprising: (a) a solid substrate comprising a surface, wherein said surface comprises a plurality of discrete regions defined on said surface,wherein said discrete regions are arranged on the surface in a regular array and are separated by inter-regional areas;(b) a population of concatemers randomly disposed on said plurality of discrete regions such that more than 95% of said plurality of discrete regions comprises a single concatemer attached thereto and said inter-regional areas are substantially free of concatemers, wherein said concatemers are generated from a circular template and comprise a plurality of monomeric units, wherein each monomeric unit comprises: (i) a target sequence of a target polynucleotide, and(ii) an adaptor,and wherein said population of concatemers comprises a plurality of different target sequences. 2. The array of claim 1, wherein said discrete regions are positively charged so as to bind said concatemers. 3. The array of claim 2, wherein said concatemers have a diameter that is approximately equal to that of the discrete regions. 4. The array of claim 1, wherein said concatemers are non-covalently immobilized to said discrete regions. 5. The array of claim 1, wherein said inter-regional areas are hydrophobic. 6. The array of claim 1, wherein the regular array comprises a rectilinear pattern. 7. The array of claim 1, wherein said discrete regions comprise capture oligonucleotides. 8. The array of claim 1, wherein said discrete regions comprise an amino group. 9. The array of claim 1, wherein the target polynucleotide is human genomic DNA. 10. The array of claim 1, wherein said discrete regions are 100 to 300 nm in diameter. 11. The array of claim 1, wherein the concatemers randomly disposed on said discrete regions are attached to a positively charged surface that binds negatively charged DNA. 12. An array comprising: (a) a solid substrate comprising a surface, wherein said surface comprises a plurality of discrete regions defined on said surface separated by inter-regional areas;(b) a population of concatemers randomly disposed on the plurality of discrete regions such that more than 95% of said plurality of discrete regions comprise a single concatemer attached thereto and said inter-regional areas are substantially free of concatemers;wherein said concatemers randomly disposed on said discrete regions are bound to a positively charged surface that binds negatively charged DNA;wherein the discrete regions are arranged on the surface as a regular array;wherein each said concatemer comprises a plurality of monomeric units, each unit comprising a target sequence of a target polynucleotide; andwherein the population of concatemers comprises a plurality of different target sequences. 13. The array of claim 12 wherein said concatemers have a diameter that is approximately equal to that of the discrete regions. 14. The array of claim 12, wherein said inter-regional areas are hydrophobic. 15. The array of claim 12, wherein the target polynucleotide is human genomic DNA. 16. The array of claim 12, wherein said discrete regions are 100 to 300 nm in diameter. 17. The array of claim 12, wherein each concatemer contains a plurality of monomeric units, each comprising: (i) a target sequence of a target polynucleotide, and(ii) an adaptor. 18. The array of claim 1, wherein said concatemers have a diameter that is approximately equal to that of the discrete regions. 19. An array comprising: (a) a solid substrate comprising a surface, wherein said surface comprises a plurality of discrete regions defined on said surface,wherein said discrete regions are arranged on the surface in a regular array and are separated by inter-regional areas;(b) a population of concatemers randomly disposed on said plurality of discrete regions such that at least a majority of said plurality of discrete regions comprises a single concatemer attached thereto and said inter-regional areas are substantially free of concatemers, wherein said concatemers are generated from a circular template and comprise a plurality of monomeric units, wherein each monomeric unit comprises: (i) a target sequence of a target polynucleotide, and(ii) an adaptor,wherein said discrete regions (i) have a diameter that is approximately equal to that of the concatemers, or(ii) have a diameter of 100 to 300 nmand wherein said population of concatemers comprises a plurality of different target sequences. 20. The array of claim 19, wherein said discrete regions are positively charged so as to bind said concatemers. 21. The array of claim 19, wherein said concatemers are non-covalently immobilized to said discrete regions. 22. The array of claim 19, wherein said inter-regional areas are hydrophobic. 23. The array of claim 19, wherein the regular array comprises a rectilinear pattern. 24. The array of claim 19, wherein more than 80% of said discrete regions have a single concatemer attached thereto. 25. The array of claim 19, wherein said discrete regions comprise capture oligonucleotides. 26. The array of claim 19, wherein said discrete regions comprise an amino group. 27. The array of claim 19, wherein the target polynucleotide is human genomic DNA. 28. The array of claim 19, wherein the concatemers randomly disposed on said discrete regions are attached to a positively charged surface that binds negatively charged DNA. 29. An array comprising: (a) a solid substrate comprising a surface, wherein said surface comprises a plurality of discrete regions defined on said surface separated by inter-regional areas;(b) a population of concatemers randomly disposed on the plurality of discrete regions such that more than 80% of said plurality of discrete regions comprise a single concatemer attached thereto and said inter-regional areas are substantially free of concatemers;wherein said concatemers randomly disposed on said discrete regions are bound to a positively charged surface that binds negatively charged DNA;wherein the discrete regions are arranged on the surface as a regular array;wherein each said concatemer comprises a plurality of monomeric units, each unit comprising a target sequence of a target polynucleotide;wherein said discrete regions (i) have a diameter that is approximately equal to that of the concatemers, or(ii) have a diameter of 100 to 300 nm, andwherein the population of concatemers comprises a plurality of different target sequences. 30. The array of claim 29, wherein said inter-regional areas are hydrophobic. 31. The array of claim 29, wherein more than 95% of the discrete regions have a single concatemer attached thereto. 32. The array of claim 29, wherein the target polynucleotide is human genomic DNA. 33. The array of claim 29, wherein each concatemer contains a plurality of monomeric units, each comprising: (i) a target sequence of a target polynucleotide, and(ii) an adaptor. 34. The array of claim 12, wherein said discrete regions comprise an amino group. 35. The array of claim 29, wherein said discrete regions comprise an amino group. 36. The array of any one of claim 1, 12, 19 or 29 wherein the target polynucleotide comprised human genomic DNA.
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