Nucleic acid concatenation
First Claim
1. A method of length-controlled concatenating nucleotide fragments, the method comprising:
- (a) providing at least two nucleotide fragments, wherein each fragment has one ligatable end and one non-ligatable end; and
(b) allowing the two fragments to ligate at the ligatable ends to form at least one oligonucleotide comprising at least two concatenated nucleotide fragments.
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Abstract
The present invention provides a method of manipulating nucleic acids. In particular, of length-controlled concatenating of nucleotide fragments, the method comprising: (a) providing at least two nucleotide fragments, wherein each fragment has one ligatable end and one non-ligatable end; and (b) allowing the two fragments to ligate at the ligatable ends to form an oligonucleotide comprising at least two concatenated nucleotide fragments. The present invention also provides an isolated oligonucleotide comprising at least two nucleotide fragments, wherein each fragment has at least one ligatable end and and one non-ligatable end, and the fragments are ligated at the ligatable ends to form the oligonucleotide.
24 Citations
36 Claims
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1. A method of length-controlled concatenating nucleotide fragments, the method comprising:
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(a) providing at least two nucleotide fragments, wherein each fragment has one ligatable end and one non-ligatable end; and (b) allowing the two fragments to ligate at the ligatable ends to form at least one oligonucleotide comprising at least two concatenated nucleotide fragments. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. A method of length-controlled concatenating nucleotide fragments, the method comprising the steps of:
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(a) providing at least one oligonucleotide comprising at least one nucleotide fragment, wherein the oligonucleotide has ligatable ends; (b) allowing the at least one oligonucleotide to self-circularize at its ligatable ends; (c) selecting the at least one self-circularized oligonucleotide; (d) treating the selected circularized oligonucleotide with at least one restriction enzyme to obtain at least one oligonucleotide with one ligatable end and one non-ligatable end; and (e) concatenating at least two oligonucleotides at the ligatable ends to form a concatenated oligonucleotide comprising at least two nucleotide fragments. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31)
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- 32. An isolated oligonucleotide comprising at least two nucleotide fragments, wherein each fragment has at least one ligatable end and and one non-ligatable end, and the fragments are ligated at the ligatable ends to form the oligonucleotide.
Specification