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Application of graphene oxide

A technology of graphene and stone oxide, which is applied in the application field of graphene oxide, can solve the problem of unsatisfactory comprehensive optimization effect of carbon nanomaterials, and achieve the effect of eliminating amplification primer dimer, low cost, and excellent comprehensive effect

Inactive Publication Date: 2014-08-27
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The technical problem to be solved by the present invention is to provide a new application of graphene oxide in PCR technology in order to overcome the defect that the comprehensive optimization effect of carbon nanomaterials that has been applied to PCR technology is not ideal.

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  • Application of graphene oxide
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  • Application of graphene oxide

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0027] Reference Example 1 Preparation of Graphene Oxide A

[0028] The preparation method and structural information characterization of graphene oxide A are as described in the literature (Reduction of graphene oxide via L-ascorbic acid.Chem.Commun., 2010, 46, 1112-1114), specifically for first using KMnO 4 / H 2 SO 4 Oxidize graphite powder to obtain oxidized graphite, and then undergo ultrasonic treatment to obtain a sheet of graphene oxide exfoliated therefrom. GO was observed by atomic force microscopy (AFM) and transmission electron microscopy (TEM), and its height was 1.2 nm, consistent with the known height of monolayer GO.

[0029]The size and distribution statistics of graphene oxide A sheets are as follows: a large sheet is a single GO sheet whose upper surface area is greater than 1×10 5 nm 2 , the ratio of the sum of the area of ​​large sheets of GO meeting this condition to the total upper surface area is 83.2%, and the number of large sheets meeting this con...

Embodiment 1

[0034] The influence of the graphene oxide (GO) of embodiment 1 different concentrations on the β-actin gene amplification of people

[0035] 1. Purpose of the experiment

[0036] Using the human genome as a template, amplify the 408bp β-actin gene (Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science, 1985, 230, 1350-1354.), compared with no GO , and the effect of adding different amounts of GO.

[0037] 2. Experimental methods and materials

[0038] Primer 1: 5′-GAAGAGCCAAGGACAGGTAC-3′

[0039] Primer 2: 5′-GGAAAATAGACCAATAGGCAG-3′

[0040] Composition of PCR solution system (25μl):

[0041] components

[0042] PCR steps and conditions: (1) 94°C preheating for 5 minutes, (2) 94°C denaturation for 30 seconds, (3) 55°C annealing for 40 seconds, (4) 72°C extension for 40 seconds, (5) 72°C extension for 5 minutes ; Steps (2)-(4) are repeated for 35 cycles.

[0043] 3. Experimental resu...

Embodiment 2

[0049] Example 2 PCR optimization effect of graphene oxide (GO) on different concentrations of human genome template

[0050] 1. Purpose of the experiment

[0051] Using the human genome as a template, the 408bp β-actin gene was amplified, and the PCR amplification effects before and after adding GO to different concentrations of the human genome template were compared.

[0052] 2. Experimental methods and materials

[0053] GO is GO of Reference Example 1 and Reference Example 3.

[0054] The experiment was divided into three groups, one group without GO, and the other two groups with 0.8ng / μl graphene oxide A and graphene oxide C added to the PCR solution respectively. In each group line, 5 groups of experiments were carried out, and the concentrations of the human genome template were 2, 1, 0.1, 0.01 and 0.001 ng respectively. All the other components in the PCR solution system are the same as in Example 1.

[0055] The PCR conditions are the same as in Example 1.

[0...

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Abstract

The invention discloses application of graphene oxide to polymerase chain reaction (PCR). The application is implemented according to the method that the PCR is carried out under the action of the graphene oxide. The invention finds that the graphene oxide has the advantages of obviously increasing the specificity and the sensitivity of PCR and amplifying the yield by being applied to the PCR technology for the first time and can eliminate a primer dimer formed in amplification, in addition, the graphene oxide has a wide optimization interval and can be widely applied to DNA templates of various concentrations and complex degrees. Compared with other carbon nanomaterials applied to the PCR technology, the graphene oxide has a more excellent comprehensive effect on optimizing the PCR. In addition, the graphene oxide is low in cost, mature in preparation process, and simple to operate when applied to the PCR technology.

Description

technical field [0001] The present invention relates to a new application of graphene oxide in polymerase chain reaction. Background technique [0002] PCR (polymerase chain reaction) is a technique for mass replication of DNA in vitro, which is widely used in genetic testing, medical diagnosis and other fields. With the increasing application of PCR technology, researchers continue to develop, optimize and improve PCR technology. For example, nano-PCR technology is a new technology that utilizes the unique surface physical and chemical properties and large surface area to volume ratio of nanomaterials to optimize the effect of DNA amplification in vitro. In recent years, many nanomaterials have been used for the optimization of PCR technology, mainly including: (1) metal nanoparticles, such as nano-gold or nano-silver, which can improve the specificity, reaction sensitivity and reaction speed of PCR, etc.; (2) ) semiconductor materials, such as CdSe quantum dots, can impr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/34C12Q1/68
Inventor 米丽娟张益郭守武胡钧
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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