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Plasmid for preparing DNA Marker, and construction method and application thereof

A plasmid and fragment technology, applied in the fields of molecular biology and genetic engineering, can solve the problems of complex carrier molecule construction, difficulty in ensuring batch repeatability, and high overall cost, and achieve the effect of simplifying the process

Inactive Publication Date: 2012-02-01
生工生物工程(上海)股份有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] To sum up, in the current DNA Marker production process, although the PCR amplification method mainly used is simple, it is a labor-intensive method, and it is difficult to ensure the repeatability between batches, so it is difficult to achieve large-scale production. costly method
Enzyme digestion of artificially constructed DNA molecules in the enzyme digestion method is the mainstream of future technology development, but there are still problems in the early stage of carrier molecule construction, poor flexibility in preparing different types of Markers, and incompletely uniform band brightness in the later stage of preparation. bottleneck

Method used

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  • Plasmid for preparing DNA Marker, and construction method and application thereof
  • Plasmid for preparing DNA Marker, and construction method and application thereof
  • Plasmid for preparing DNA Marker, and construction method and application thereof

Examples

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Embodiment 1

[0075] Preparation of a universal low molecular weight DNA marker

[0076] This example describes the preparation of a universal low molecular weight DNA Marker. The Marker contains 6 DNA bands, respectively 2000bp, 1000bp, 750bp, 500bp, 250bp, and 100bp. It is currently the most widely used DNA in various laboratories. Marker, for easy identification, we have enhanced the brightness of the 750bp band.

[0077] Unless otherwise specified, the biochemical reagents and PCR or nucleic acid extraction and purification kits used in this example are from Shenggong Bioengineering (Shanghai) Co., Ltd.; all primers are synthesized by Shenggong Bioengineering (Shanghai) Co., Ltd. Synthesized by solid-phase phosphoramidite triester method and purified by polyacrylamide gel electrophoresis; all DNA sequences were determined by Shenggong Bioengineering (Shanghai) Co., Ltd. by dideoxy chain termination method combined with Applied Biosystems 3730 / 3730xl DNA Analyzers Instrumental analysis; all ...

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Abstract

The invention provides a plasmid for preparing a DNA Marker, and a construction method and application thereof. The plasmid comprises different DNA fragments forming the target DNA marker; the copy number of the different DNA Marker fragments in the plasmid is more than 1, and the same single restriction enzyme cutting site is positioned between the DNA Marker fragments. The invention also provides a method for quickly preparing the DNA marker at low cost. The DNA marker prepared by the method has the characteristics of sharp band, uniform brightness or random controllability, high repeatability between batches and the like.

Description

Technical field [0001] The invention belongs to the field of molecular biology and genetic engineering, and relates to a method for producing DNA markers that are widely used in genetic engineering. Specifically, the present invention relates to a plasmid for low-cost and rapid production of high-quality DNA Marker and a construction method thereof, and also to a method and application of using the plasmid to produce a DNA Marker. Background technique [0002] Genetic engineering, also known as recombinant DNA technology, refers to a series of operations such as cutting, joining, and modifying DNA at the molecular level in vitro. It was developed on the basis of molecular biology and molecular genetics. It was born in 20 A brand-new biological science and technology in the 1970s. Although this technology is a young technology, it has played a very important role in the research and application of life sciences, biomedicine, and even physics and polymer science. [0003] In the in...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/70C12N15/66C12N15/11C12N15/10C12R1/19
Inventor 李威万军飞
Owner 生工生物工程(上海)股份有限公司
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