Kits for sorting and identifying polynucleotides
First Claim
1. A kit for sorting and identifying polynucleotides, the kit comprising:
- one or more solid phase supports each having one or more spatially discrete regions, each such region having a uniform population of substantially identical tag complements covalently attached, and the tag complements each being selected from a single minimally cross-hybridizing set of oligonucleotides having a length between 10 and 30 nucleotides, wherein each oligonucleotide of the set is separately synthesized and differs from every other oligonucleotide of the set by at least three nucleotides.
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Abstract
The invention provides a method of tracking, identifying, and/or sorting classes or subpopulations of molecules by the use of oligonucleotide tags. Oligonucleotide tags of the invention comprise oligonucleotides selected from a minimally cross-hybridizing set. Preferably, such oligonucleotides each consist of a plurality of subunits 3 to 9 nucleotides in length. A subunit of a minimally cross-hybridizing set forms a duplex or triplex having two or more mismatches with the complement of any other subunit of the same set. The number of oligonucleotide tags available in a particular embodiment depends on the number of subunits per tag and on the length of the subunit.
An important aspect of the invention is the use of the oligonucleotide tags for sorting polynucleotides by specifically hybridizing tags attached to the polynucleotides to their complements on solid phase supports. This embodiment provides a readily automated system for manipulating and sorting polynucleotides, particularly useful in large-scale parallel operations, such as large-scale DNA sequencing, mRNA fingerprinting, and the like, wherein many target polynucleotides or many segments of a single target polynucleotide are sequenced simultaneously.
244 Citations
16 Claims
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1. A kit for sorting and identifying polynucleotides, the kit comprising:
one or more solid phase supports each having one or more spatially discrete regions, each such region having a uniform population of substantially identical tag complements covalently attached, and the tag complements each being selected from a single minimally cross-hybridizing set of oligonucleotides having a length between 10 and 30 nucleotides, wherein each oligonucleotide of the set is separately synthesized and differs from every other oligonucleotide of the set by at least three nucleotides. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
Specification