Invention Application
- Patent Title: Length determination of nucleic acid repeat sequences by discontinuous primer extension
- Patent Title (中): 通过不连续引物延伸确定核酸重复序列的长度
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Application No.: US11731192Application Date: 2007-03-30
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Publication No.: US20070178519A1Publication Date: 2007-08-02
- Inventor: Kenneth Livak , Adam Lowe , Andrew Blasband
- Applicant: Kenneth Livak , Adam Lowe , Andrew Blasband
- Applicant Address: US CA Foster City
- Assignee: Applera Corporation
- Current Assignee: Applera Corporation
- Current Assignee Address: US CA Foster City
- Main IPC: C12Q1/68
- IPC: C12Q1/68 ; C12P19/34 ; C07H21/04

Abstract:
Disclosed is a method for determining the number of repeat units in a repeat region of a target nucleic acid. In a first aspect, the method of the invention includes the steps of annealing a primer to a target nucleic acid; performing a first primer extension reaction using a first primer extension reagent; separating the target-primer hybrid and unreacted first primer extension reagent; performing a second primer extension reaction using a second primer extension reagent, wherein at least one of the first or second primer extension reagents includes an extendible nucleotide having a label attached thereto; separating the target-primer hybrid from unreacted second primer extension reagent; measuring a signal produced by the label; treating the label so as to render the label undetectable; and repeating the above steps until the signal is substantially less than a signal detected in a previous cycle. In a second aspect, the method of the invention includes the steps of annealing a primer to a target nucleic acid; performing a first primer extension reaction using a first primer-extension reagent; separating the target-primer hybrid from unreacted first primer extension reagent; performing a second primer extension reaction using a second primer extension reagent and with a primer termination reagent, the primer termination reagent including a nucleotide terminator having a label attached thereto; separating the target-primer hybrid from unreacted second primer extension reagent and unreacted primer termination reagent; measuring a signal produced by the label; and repeating the above steps until a signal is detected indicating incorporation of the nucleotide terminator. The invention further includes kits useful for practicing the above methods.
Public/Granted literature
- US07414115B2 Length determination of nucleic acid repeat sequences by discontinuous primer extension Public/Granted day:2008-08-19
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