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Method for optimizing PCR with composite nano material

A technology of nano-materials and nano-metals, which is applied in the field of combined nano-materials to optimize PCR, can solve problems such as effect differences, and achieve low application costs, obvious effects, and easy-to-master effects

Inactive Publication Date: 2010-08-04
山东大正医疗器械股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] After comparison, the above two patents are essentially different from this patent, and the effects achieved are significantly different

Method used

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  • Method for optimizing PCR with composite nano material
  • Method for optimizing PCR with composite nano material
  • Method for optimizing PCR with composite nano material

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0032] 1. Preparation of nanomaterial suspension solution:

[0033] Commercially available products can be used as solid powder or colloid of nanomaterials. Taking the nano-carbon powder suspension solution with a concentration of (W / V) 10mg / mL as an example, first accurately weigh 10mg of sterilized nano-carbon powder and place it in a sterilized Slowly add sterilized water to a small 1.5mL centrifuge tube with a pipette until the volume is 1mL; then close the cap of the centrifuge tube tightly and place it in an ultrasonic oscillator for ultrasonic treatment for a certain period of time. The specific ultrasonic power and ultrasonic time should be expressed in nanometers. It is better that the carbon powder is completely suspended and can be placed at room temperature for at least 10 minutes without accumulation and precipitation.

[0034] 2. Preparation of composite nanomaterial suspension solution:

[0035] The nanomaterial suspension and organic reagents are combined and ...

Embodiment 1

[0054] A method for combining nanomaterials to optimize PCR, the steps are as follows:

[0055] Combined nano material B11 (nano carbon powder (<30nm, final concentration 1.3ug / ul) + ethylene glycol (final concentration 13% V / V)) was used to optimize the amplified DNA fragment with a fragment length of about 700bp.

[0056] 1. Configuration of PCR reaction system:

[0057] Configure the system in thin-walled PCR tubes in the following order and dosage:

[0058] 10×PCR buffer 1.5μL

[0059] dNTP substrate (2.5mM) 1.2μL

[0060] Primer a (2.0μM) 1μL

[0061] Primer b (2.0μM) 1μL

[0062] Template (0.1μg / μL) 0.3μL

[0063] MgCl 2 (25mM) 1.2μL

[0064] Taq enzyme (5U / μL) 0.2μL

[0065] Combined nanomaterial suspension 6 μL

[0066] Sterilized pure water 2.6μL

[0067] Wherein, the template is human genome DNA, and Taq enzyme is purchased from Shanghai Sangong (Sangon).

[0068] A total of 6 tubes of PCR system (1, 2, 3, 1', 2', 3') were configured.

[0069]

[0070]...

Embodiment 2

[0077] A method for combining nanomaterials to optimize PCR, the steps are as follows:

[0078] Combined nanomaterial B17 (multi-walled carbon nanotubes (final concentration 1.3ug / ul) + 1,2-propanediol (final concentration 13% V / V)) optimizes the results of amplifying a DNA fragment with a fragment length of about 700bp.

[0079] 1. Configuration of PCR reaction system:

[0080] Configure the system in thin-walled PCR tubes in the following order and dosage:

[0081] 10×PCR buffer 1.5μL

[0082] dNTP substrate (2.5mM) 1.2μL

[0083] Primer a (2.0μM) 1μL

[0084] Primer b (2.0μM) 1μL

[0085] Template (0.1μg / μL) 0.3μL

[0086] MgCl 2 (25mM) 1.2μL

[0087] Taq enzyme (5U / μL) 0.2μL

[0088] Combined nanomaterial suspension 6 μL

[0089] Sterilized pure water 2.6μL

[0090] Wherein, the template is human genome DNA, and Taq enzyme is purchased from Shanghai Sangong (Sangon).

[0091] A total of 6 tubes of PCR system (1, 2, 3, 1', 2', 3') were configured.

[0092]

...

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Abstract

The invention relates to a method for optimizing PCR with composite nano material. The method includes the following steps: (1) preparing the composite nano material; (2) configuring a PCR system; (3) adding the composite nano material into the PCR system; (4) setting and operating PCR conditions; and (5) testing PCR product; and the composite nano material comprises nano material and organic reagent. Compared with the prior part, the method as a new outcome integrated with the nano technology and the biotechnology has the following advantages that: by adding a certain amount of nano material and organic reagent in the amplification system, the method achieves the specificity of amplified target gene, the effect is remarkable, moreover, the application cost is low, use is easy, and the method is easy to master.

Description

Technical field: [0001] The invention relates to the field of biotechnology, in particular to a method for optimizing PCR by combining nanomaterials. Background technique: [0002] PCR is polymerase chain reaction, also known as in vitro enzymatic gene amplification. It is a gene amplification technology that simulates DNA replication in vitro. This technology was invented by K. Mullis in 1985. Under the guidance of specific primers and the role of specific primers, a large number of genes can be replicated in a test tube in a short period of time. Generally speaking, the target gene can be amplified to a million times within a few hours. [0003] Over the past 20 years, PCR technology has gradually matured through continuous progress and development, forming a complete technical system. Conventional PCR technology has been widely used in the research of genetics, microbiology and even the entire life science field due to its simple operation and high success rate. At pres...

Claims

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Application Information

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IPC IPC(8): C12P19/34
Inventor 张治洲
Owner 山东大正医疗器械股份有限公司
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