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Small-DNA-molecular-weight standard, and standard plasmid and preparation method thereof

A technology of molecular weight standards and molecular weight, applied in biochemical equipment and methods, recombinant DNA technology, DNA/RNA fragments, etc., can solve problems such as large demand

Active Publication Date: 2016-10-12
北京博迈德基因工程技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the length of the PCR product of PCR amplification for identification is usually between 100-600bp, and the demand is large
However, because the DNA fragments are too small, commercially available plasmid enzyme-cut small molecule DNA molecular weight markers less than 700bp in size are rare on the market, and there is no molecular weight marker that constructs all 7 fragments on a single plasmid
The construction method of this plasmid has not been reported in professional journals and patents at home and abroad

Method used

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  • Small-DNA-molecular-weight standard, and standard plasmid and preparation method thereof
  • Small-DNA-molecular-weight standard, and standard plasmid and preparation method thereof
  • Small-DNA-molecular-weight standard, and standard plasmid and preparation method thereof

Examples

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Effect test

Embodiment 1

[0116] A small deoxyribonucleic acid molecular weight standard, the small deoxyribonucleic acid molecular weight standard is a small deoxyribonucleic acid sequence, and the small deoxyribonucleic acid sequence is 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 100bp fragments, 1 or 2 or 3 200bp fragments, 1 or 2 300bp fragments, 1 or 2 400bp fragments , a 500bp fragment, a 600bp fragment and a 700bp fragment, all of the above small fragments or small DNA sequences are constructed on the same vector.

Embodiment 2

[0118] A small deoxyribonucleic acid molecular weight standard, the small deoxyribonucleic acid molecular weight standard is a small deoxyribonucleic acid sequence, and the small deoxyribonucleic acid sequence is 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 100bp fragments, 1 or 2 or 3 200bp fragments, 1 or 2 300bp fragments, 1 or 2 400bp fragments , a 500bp fragment, a 600bp fragment, a 700bp fragment and a 800bp fragment, all of the above small fragments or small deoxyribonucleic acid sequences are constructed on the same vector.

Embodiment 3

[0120] A small deoxyribonucleic acid molecular weight standard, the small deoxyribonucleic acid molecular weight standard is a small deoxyribonucleic acid sequence, and the small deoxyribonucleic acid sequence is 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 100bp fragments, 1 or 2 or 3 200bp fragments, 1 or 2 300bp fragments, 1 or 2 400bp fragments , 1 fragment of 500bp, 1 fragment of 600bp, 1 fragment of 700bp, 1 fragment of 800bp and 1 fragment of 900bp, all of the above small fragments or small DNA sequences are constructed on the same vector .

[0121] In a further preferred embodiment, the vector described in Examples 1-3 is a plasmid, more preferably a plasmid PUC19.

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Abstract

The invention relates to a small-DNA-molecular-weight standard, and a standard plasmid and a preparation method thereof. Small DNA sequences with the length of 100 bp, 200 bp, 300 bp, 400 bp, 500 bp, 600 bp, 700 bp and the like are constructed onto a plasmid by using overlapping polymerase chain reaction, restriction endonuclease digestion, DNA connection and other methods. After the plasmid is completely digested by single restriction enzyme, and seven uniform-brightness strips can be obtained through enzyme digestion. The small-DNA-molecular-weight standard is used for preparing a DNA standard reference substance, and is used for indicating the relative value of DNA molecular weight in agarose electrophoresis.

Description

technical field [0001] The invention relates to an indispensable electrophoresis consumable in the fields of molecular biology and genetic engineering, in particular to a small deoxyribonucleic acid (DNA) molecular weight standard substance, a small deoxyribonucleic acid molecular weight standard substance particle and a preparation method thereof . Background technique [0002] DNA molecular weight standard is a reagent used to indicate the relative size of DNA's swimming speed in agarose electrophoresis, which is a mixture of multiple DNA fragments of known specific sizes. At present, the main preparation methods include PCR amplification method, plasmid digestion method or a combination of the two methods. The PCR method is to design primers, obtain DNA fragments of a specific size by in vitro PCR amplification, and then purify and quantify them, and then mix them together according to a certain mass ratio. The enzyme digestion method is to clone fragments of different ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/66
CPCC12N15/11C12N15/66C12N2310/10C12N2330/50
Inventor 齐建国吴立群
Owner 北京博迈德基因工程技术有限公司
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