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Optimized method for improving polymerase chain reaction (PCR) expanding effect by nano carbon powder

A nano-carbon powder and technology of optimization method are applied in the field of polymerase chain reaction amplification, which can solve the problems of high price and limited scope of wide application, and achieve the effects of wide price, obvious effect and easy separation.

Inactive Publication Date: 2008-03-05
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether it is single-walled carbon nanotubes or colloidal gold, due to their high technical requirements for production and preparation, they are expensive and their wide range of applications is limited.

Method used

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  • Optimized method for improving polymerase chain reaction (PCR) expanding effect by nano carbon powder
  • Optimized method for improving polymerase chain reaction (PCR) expanding effect by nano carbon powder
  • Optimized method for improving polymerase chain reaction (PCR) expanding effect by nano carbon powder

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Experimental program
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Effect test

preparation example Construction

[0029] 2. Preparation of nano carbon powder suspension solution:

[0030] Nano-carbon powder solid powder can be commercially available products, which belongs to the prior art and will not be described in detail. Take 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 1.5mL small centrifuge tube, and pipette Add sterilized water slowly to a volume of 1 mL. Then close the cap of the centrifuge tube 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 based on the fact that the nano-carbon powder is completely suspended and can be placed at room temperature for at least 10 minutes without accumulation and precipitation.

[0031] 3. Configuration of PCR system:

[0032] According to the existing technology, the PCR system is different accordin...

Embodiment 1

[0049] Nano carbon powder optimizes the amplified DNA fragment with a fragment length of 12kb

[0050] 1. Configuration of PCR reaction system:

[0051] Configure the system in a thin-walled PCR tube in the following order and dosage.

[0052] 10×PCR buffer 2.5μL

[0053] dNTP substrate (2.5mM) 5μL

[0054] Primer 1 (4.0μM) 1.9μL

[0055] Primer 2 (4.0μM) 1.9μL

[0056] Template (0.1μg / μL) 0.5μL

[0057] Mg 2+ (25mM) 2.8μL

[0058] Taq enzyme (5U / μL) 0.5μL

[0059] Pfu enzyme (2.5U / μL) 0.5μL

[0060] Among them, the template is λDNA, which was purchased from Canada BBI Company, and Taq enzyme and Pfu enzyme were purchased from Beijing Sanbo Yuanzhi Bioengineering Company.

[0061] A total of 5 PCR system tubes were configured, numbered from 0 to 4.

[0062] 2. Add 0 μL, 1.0 μL, 2.0 μL, 3.0 μL, and 4.0 μL of nano-carbon powder suspension solution in sequence from tube 0 to tube 5. Finally, the total volume was made up to 25 μL with double distilled water.

[0063] 3...

Embodiment 2

[0071] Nano carbon powder optimizes the amplified DNA fragment with a fragment length of 15kb

[0072] 1. Configuration of PCR reaction system:

[0073] Configure the system in a thin-walled PCR tube in the following order and dosage.

[0074] 10×PCR buffer 2.5μL

[0075] dNTP substrate (2.5mM) 5μL

[0076] Primer 1 (4.0μM) 1.9μL

[0077] Primer 2 (4.0μM) 1.9μL

[0078] Template (0.1μg / μL) 0.5μL

[0079] Mg 2+ (25mM) 2.8μL

[0080] Taq enzyme (5U / μL) 0.5μL

[0081] Pfu enzyme (2.5U / μL) 0.5μL

[0082] Among them, the template is λDNA, which was purchased from Canada BBI Company, and Taq enzyme and Pfu enzyme were purchased from Beijing Sanbo Yuanzhi Bioengineering Company.

[0083] A total of 5 PCR system tubes were configured, numbered from 0 to 4.

[0084] 2. Add 0 μL, 1.0 μL, 2.0 μL, 3.0 μL, and 4.0 μL of nano-carbon powder suspension solution in sequence from tube 0 to tube 5. Finally, the total volume was made up to 25 μL with double distilled water.

[0085] 3...

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Abstract

The present invention is one optimized method of raising the effect of PCR amplification with nanometer carbon powder, and belongs to the field of biological technology. The method is to add certain amount of nanometer carbon powder into the PCR amplification system on the basis of available PCR amplification method. The present invention has obvious optimizing effect, simple operation, and easy separation of the added matter from the system for convenient post application of the PCR resultant.

Description

technical field [0001] The invention relates to a polymerase chain reaction (PCR) amplification method in the field of biotechnology, in particular to an optimization method for improving the polymerase chain reaction (PCR) amplification effect by nano carbon powder. Background technique [0002] Polymerase chain reaction (PCR), also known as in vitro enzymatic gene amplification, is a gene amplification technology that simulates DNA replication in vitro. This technology was invented by K.Mullis in 1985. Under the action of enzymes and the guidance of specific primers, a large number of gene duplications that only occur when organisms cell division and proliferation can be realized in a test tube in a very short period of time. Generally speaking, within a few hours. Amplify the gene of interest to a million-fold. [0003] Over the past 20 years, PCR technology has gradually matured through continuous progress and development, forming a complete set of technical systems. C...

Claims

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Application Information

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IPC IPC(8): C12P19/34C12Q1/68
Inventor 张治洲汪名春
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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