Preparation of Nucleic Acid Templates for Solid Phase Amplification
First Claim
1. A method of preparing nucleic acid template constructs for solid-phase nucleic acid amplification which comprises performing the following ligation reactions a) and b):
- a) a ligation reaction in which the first end of one or more target polynucleotide molecules are ligated to surface-bound adaptor polynucleotide molecules; and
b) a ligation reaction in which solution-phase adaptor polynucleotide molecules are ligated to the second end of said target polynucleotide molecules;
thereby producing one or more template constructs attached to a solid support, wherein each of said template constructs comprises a target polynucleotide molecule ligated to a surface-bound adaptor polynucleotide molecule and a solution-phase polynucleotide molecule.
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Abstract
The invention relates to a method of preparing template constructs for solid-phase nucleic acid amplification and to use of the templates in methods of solid-phase nucleic acid amplification. The method involves carrying out two ligation reactions: (a) a ligation reaction in which the first end of one or more target polynucleotide molecules are ligated to surface-bound adaptor polynucleotide molecules, and (b) a ligation reaction in which solution-phase adaptor polynucleotide molecules are ligated to the second end of said target polynucleotide molecules, in order to produce one or more template constructs attached to a solid support.
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Citations
24 Claims
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1. A method of preparing nucleic acid template constructs for solid-phase nucleic acid amplification which comprises performing the following ligation reactions a) and b):
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a) a ligation reaction in which the first end of one or more target polynucleotide molecules are ligated to surface-bound adaptor polynucleotide molecules; and b) a ligation reaction in which solution-phase adaptor polynucleotide molecules are ligated to the second end of said target polynucleotide molecules; thereby producing one or more template constructs attached to a solid support, wherein each of said template constructs comprises a target polynucleotide molecule ligated to a surface-bound adaptor polynucleotide molecule and a solution-phase polynucleotide molecule. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 24)
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23. (canceled)
Specification