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GB/T 41799-2022: Assay method of restriction endonuclease impurity
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| GB/T 41799-2022 | English | 189 |
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Assay method of restriction endonuclease impurity
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GB/T 41799-2022
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PDF similar to GB/T 41799-2022
Standard similar to GB/T 41799-2022 GB/T 41907 GB/T 44829 GB/T 44828 Basic data | Standard ID | GB/T 41799-2022 (GB/T41799-2022) | | Description (Translated English) | Assay method of restriction endonuclease impurity | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | C27 | | Classification of International Standard | 07.080 | | Word Count Estimation | 10,187 | | Date of Issue | 2022-10-14 | | Date of Implementation | 2023-05-01 | | Issuing agency(ies) | State Administration for Market Regulation, China National Standardization Administration | GB/T 41799-2022: Assay method of restriction endonuclease impurity---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
Assay method of restriction endonuclease impurity
ICS 07.080
CCSC27
National Standards of People's Republic of China
Restriction endonuclease impurity detection method
Published on 2022-10-12
2023-05-01 Implementation
State Administration for Market Regulation
Released by the National Standardization Administration
directory
Preface III
Introduction IV
1 Scope 1
2 Normative references 1
3 Terms and Definitions 1
4 Principle 1
5 Reagents or materials 1
6 Instruments 2
7 Test Step 2
Appendix A (normative) agarose gel electrophoresis 4
foreword
This document is in accordance with the provisions of GB/T 1.1-2020 "Guidelines for Standardization Work Part 1.Structure and Drafting Rules of Standardization Documents"
drafted.
Please note that some content of this document may be patented. The issuing agency of this document assumes no responsibility for identifying patents.
This document is proposed and managed by the National Tool Enzyme Standardization Working Group (SAC/SWG11).
This document is drafted by. Fujian Nansheng Technology Co., Ltd., General Biological (Anhui) Co., Ltd., Xiahe (Shenzhen) Biotechnology Co., Ltd.
Co., Ltd., Xiamen Yinxiang Group Co., Ltd., Suzhou Kunqi Biotechnology Co., Ltd., Xiamen CIMC Detection Technology Co., Ltd., Shanghai Yingzi
Life Technology Co., Ltd., Leigu (Xiamen) Biopharmaceutical Co., Ltd., Beijing University of Chemical Technology, Shandong University, Angel Yeast Co., Ltd.,
Fudan University, Wuhan University of Science and Technology, Beijing Zhongtian Biaoke Standardization Technology Research Institute Co., Ltd., Xiamen Aide Biomedical Technology Co., Ltd.
company.
The main drafters of this document. Huang Facan, Yong Jingui, Zheng Dengzhong, Zhang Zhigang, Chen Jinchun, Chen Xiulan, Jiang Feng, Zhao Yi, Zhong Jiang, Zhu Li, Yao Juan,
Yang Zhonghua, Li Chao, Xu Wenlai, Wang Yongcheng.
Introduction
Restriction endonucleases are a class of DNA hydrolases that recognize specific nucleotide sequences in double-stranded DNA.
DNA, resulting in 5'-PO3 and 3'-OH ends. The substrates for restriction endonucleases are deoxyribonucleic acid (DNA), other nucleic acids
The contamination of endonuclease or exonuclease will lead to the degradation or disappearance of the substrate, and reducing the contamination of other endonucleases or exonuclease is limited.
The most important quality assurance of the manufacturing endonucleases. Formulate national standards for the detection method of restriction endonuclease impurities to promote this kind of work.
The industrialization of enzymes is of great significance for the production and use of restriction endonucleases.
Restriction endonuclease impurity detection method
1 Scope
This document describes the detection methods for impurities such as contaminating other endonucleases or exonucleases in restriction endonucleases.
This document applies to the detection of restriction endonuclease impurities.
2 Normative references
There are no normative references in this document.
3 Terms and Definitions
The following terms and definitions apply to this document.
3.1
Hydrolysis of DNA by endonucleation to generate 5'-PO3 and 3'-OH ends, identifying DNA water of specific nucleotide sequences in double-stranded DNA
lyase.
3.2
Impurity
Other endonucleases and exonucleases that affect the presence of nucleic acid substrates.
4 Principles
Since ФX174DNA has no BamHI restriction site, λDNA has 5 BamHI restriction sites, and 6 spectra are generated after complete enzymatic digestion
bands, their sizes and nucleotide sequences are known. There is no other endonuclease contamination in the sample, and the spectrum formed by the enzymatic hydrolysis product on the electrophoresis plate
The band has nothing to do with the long time of enzymolysis and the use of too much enzyme.
Since BamHI cuts the DNA double-strand to produce a pair of sticky ends, each of which has 5 bases that lose their complementary bases to form a single-strand protruding single strand.
It is susceptible to the action of exonuclease to lose several bases and even form a blunt end; if the DNA is ligated and then digested with BamHI, the result
cannot be cut. If the site is in the exon region or coding region, it means that the corresponding amino acid sequence has changed, and even some biological
traits changed. For example, the BamHI recognition site contained in plasmid pBR322 is on its anti-tetracycline gene.
Dicer-contaminated samples with excess BamHI cleaved pBR322 for a long time, and then ligated and closed the loop to re-transform the recipient bacteria.
However, pBR322 transformants without such enzyme digestion could grow. Another example is the BamHI site contained in plasmid pUC19
Spot on the α-complementary peptide encoding LacZ, if the aforementioned situation occurs, it will lead to changes in the amino acid sequence of the peptide chain, and finally
The LB medium of IPTG and X-gal changed from blue spots to white spots. If the number of leukoplakia accounted for more than 10% of the total number of plaques, it was judged
The sample was determined to be contaminated with exonuclease.
5 Reagents or materials
5.1 lambda phage deoxyribonucleic acid (lambda DNA).
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