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Suturing Toehold activation method used for controlling DNA strand displacement reaction and toolkit

A technology of displacement reaction and DNA chain, applied in the field of molecular biology, can solve the problems of limiting the design flexibility of dynamic DNA nanodevices

Active Publication Date: 2015-03-11
SHANDONG UNIV
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Problems solved by technology

However, the above methods all need to design the recognition site or sequence of the stimulus in the toehold part, which greatly limits the flexibility of dynamic DNA nanodevices design

Method used

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  • Suturing Toehold activation method used for controlling DNA strand displacement reaction and toolkit
  • Suturing Toehold activation method used for controlling DNA strand displacement reaction and toolkit
  • Suturing Toehold activation method used for controlling DNA strand displacement reaction and toolkit

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Embodiment 1

[0027] Experimental part of embodiment 1

[0028] 1.1 Materials and Instruments

[0029] All DNA sequences were synthesized by Shanghai Bioengineering Co., Ltd. (China). Mercury nitrate dissolved in 2-5% nitric acid was purchased from Bailingwei Chemical Technology Co., Ltd. (China). ATP, CTP, GTP, UTP and 40% acrylamide, ammonium persulfate, TEMED were all purchased from Shanghai Bioengineering Co., Ltd. (China). All reagents were of analytical grade, and the water used was ultrapure water. All fluorescence experiments were performed on a Japanese Hitachi FL-7000 fluorescence spectrophotometer. Spectral conditions: the excitation voltage is 700V, the excitation wavelength and emission wavelength are 495nm and 530nm, respectively, and the slit width is 10nm. 1.2 Probe Preparation for Binding-Induced Suture Toehold Strand Displacement Reaction

[0030] To prepare Hg 2+ To induce suture RS in Toehold-mediated strand displacement reaction, add 10 μL, 10 μM FAM-labeled TB an...

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Abstract

The invention provides a suturing Toehold activation method used for controlling a DNA strand displacement reaction and a toolkit. A novel Toehold activation method is developed based on hairpin reconstruction caused by mutual action of a DNA hairpin structure and different environmental irritants (Hg2+ or ATP). The invention develops a novel Toehold activation method. The extra level of the DNA strand displacement reaction can be accurately controlled by changing the concentration of the environmental irritants.

Description

Technical field: [0001] The invention belongs to the field of molecular biology, and in particular relates to a suture Toehold activation method and toolkit for regulated DNA strand displacement reactions. Background technique: [0002] Due to the specificity and predictability of the Watson-Crick base pairing principle, DNA has become a material motif for building structures at the nanometer scale 1,2 . With the progress of dynamic DNA nanotechnology, a series of dynamic DNA nanodevices have been successfully constructed, such as nanomachines 3,4 , logic gate 5,6 , catalytic amplifier 7,8 Wait. Most of the design and construction of functionalized and automated nanodevices are based on Toehold-mediated strand displacement reactions. Toehold-mediated strand displacement reaction uses prehybridized double-stranded DNA as the starting point of the reaction, uses the single-stranded region (toehold) of the trigger strand to trigger the strand displacement reaction, and fin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 姜玮王磊朱静
Owner SHANDONG UNIV
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