Methods of selecting and amplifying polynucleotides
A technology of polynucleotides and template polynucleotides, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problems of increased sample processing time, loss of template samples, high clustering density, etc.
Active Publication Date: 2018-04-20
ILLUMINA INC
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However, such titrations can result in loss of valuable flow cell channels due to cluster densities that are too high or too low, template sample loss, increased reagent volumes required, or increased sample processing time
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[0022] In certain embodiments, the invention relates to methods of selecting and controlling the density of different molecular species derivatized on a surface. In specific embodiments, the molecular species are nucleic acids with distinct sequences. The invention is particularly useful for controlling the density of nucleic acid clusters produced on solid supports. An advantage of these methods is that they reduce or even eliminate the need for multiple sample titration steps to control the density of molecules on the surface. Another advantage of the present invention is the ability to select a portion of a nucleic acid sample by sequence selective hybridization to capture oligonucleotides.
[0023] The methods set forth herein can be used in conjunction with the methods described in US application 12 / 395229, including methods for controlling cluster density, for example, through the use of capture oligonucleotides on solid supports. In specific embodiments, the methods s...
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The invention provides a method for selection and amplification of polynucleotides on a solid phase carrier. The method comprises the following steps of providing amplified oligonucleotides fixed to the solid phase carrier, carrying out hybridization on an oligonucleotide probe group and an amplified oligonucleotide subgroup, wherein each one of the oligonucleotide probes comprises a first part complementary with the amplified oligonucleotides and a second part from a sequence in a template polynucleotide selection zone, carrying out extension reaction so that the hybrid amplified oligonucleotides are extended and a capture oligonucleotide group bonding with carriers is produced, wherein each one of the capture oligonucleotides in the group comprises a sequence complementary with the template polynucleotide selection zone, applying the template polynucleotide group to the solid phase carrier, carrying out extension hybridization with the capture oligonucleotides of the bonding carriers of the template polynucleotides so that the extension product complementary with the template polynucleotide is produced, and carrying out amplification extension on the product to obtain a solid phase amplification product. The method can control density of different types of molecules on the solid phase carrier surface.
Description
[0001] This application is a divisional application of a Chinese invention patent application with an application date of August 25, 2009, an application number of 200980162143.X, and an invention title of "Method for Selecting and Amplifying Polynucleotides". technical field [0002] The present invention relates to the field of nucleic acid amplification. More specifically, the present embodiments provide methods for selecting one or more regions of a nucleic acid sample on a solid support and growing nucleic acid clusters directly on the solid support while eliminating the need for a multiple sample titration step. Background technique [0003] Several publications and patent documents are cited in this application to more fully describe the state of the art to which this invention pertains. The entire disclosure content of each of these publications and documents is incorporated herein by reference. [0004] Many methods for high-throughput nucleic acid sequencing rely ...
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IPC IPC(8): C12Q1/6837
Inventor 安德里亚·萨博罗伯托·里加蒂敏-瑞·理查德·沈
Owner ILLUMINA INC