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GB/T 34776-2017: T4 DNA ligase -- Detection methods of activity and impurities of enzyme
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| GB/T 34776-2017 | English | 179 |
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T4 DNA ligase -- Detection methods of activity and impurities of enzyme
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GB/T 34776-2017
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Basic data | Standard ID | GB/T 34776-2017 (GB/T34776-2017) | | Description (Translated English) | T4 DNA ligase -- Detection methods of activity and impurities of enzyme | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | A40 | | Classification of International Standard | 07.080 | | Word Count Estimation | 9,964 | | Date of Issue | 2017-11-01 | | Date of Implementation | 2018-05-01 | | Regulation (derived from) | National Standard Announcement 2017 No. 29 | | Issuing agency(ies) | General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, Standardization Administration of the People's Republic of China | GB/T 34776-2017: T4 DNA ligase -- Detection methods of activity and impurities of enzyme---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.
T4 DNA ligase-Detection methods of activity and impurities of enzyme
ICS 07.080
A40
National Standards of People's Republic of China
T4 DNA ligase activity and impurity detection methods
Posted.2017-11-01
2018-05-01 implementation
General Administration of Quality Supervision, Inspection and Quarantine of People's Republic of China
China National Standardization Administration released
Foreword
This standard was drafted in accordance with the rules given in GB/T 1.1-2009.
This standard by the National Tool Enzyme Working Group (SAC/SWG11) centralized.
This standard drafting unit. China Institute of Testing Technology, Shenzhen Huaneng Kang Gene Technology Co., Ltd., Fujian Huacan Pharmaceutical Co., Ltd.
The main drafters of this standard.Zhou Lihua, Li Huaping, Sheng Tong, Sun Dengfeng, Tan Huibiao, Huangfa Can, Jin Hong, Ma Li Xia, Ye Deping, Li Hua,
Wang Zhi, Shen Xingzhong, Tan peace.
T4 DNA ligase activity and impurity detection methods
1 Scope
This standard specifies the T4DNA ligase activity, impurity detection methods.
This standard applies to research as a molecular biology T4DNA ligase.
2 Normative references
The following documents for the application of this document is essential. For dated references, only the dated version applies to this article
Pieces. For undated references, the latest edition (including all amendments) applies to this document.
GB/T 6682 analytical laboratory water specifications and test methods
YY/T 0087 electrophoresis device
YY/T 0657 medical centrifuge
JJG646 pipette test procedures
3 Terms and definitions
The following terms and definitions apply to this document.
3.1
DNA ligase DNAligase
A gapped enzyme that blocks the DNA strand, catalyzing the 3'-hydroxylation between adjacent nucleotide molecules with the aid of the energy provided by ATP or NAD hydrolysis
The base and the 5'-phosphate group form a phosphodiester bond.
Note. DNA ligase-catalyzed substrates can be either DNA or RNA. Depending on the type of enzyme, cofactors that produce high energy intermediates in the reaction may be
ATP or NAD.
3.2
T4 DNA ligase T4 DNAligase
A T4 phage gene 30 encoding product, with ATP as a cofactor, catalyzes the double-stranded DNA or RNA 5'-phosphate end
The terminal and 3'-hydroxyl ends form phosphodiester bonds.
Note. His-Tag, with a molecular mass of approximately 70 ku, is derived from the E. coli strain transformed with T4 DNA ligase expression vector. The main role is catalysis
Linkage between DNA ends repairs single-stranded nicks in double-stranded nucleic acids.
3.3
Unit of activity
In 20μL 1 × T4 DNA ligase reaction buffer, reaction conditions at 16 ℃, 30min can make 50% Hind Ⅲ eliminate
The amount of enzyme required to ligate the [lambda] DNA fragment [5 'end of 0.12 [mu] mol/L (300 [mu] g/mL] is 1 unit of activity.
4 Abbreviations
The following abbreviations apply to this document.
PCR polymerase chain reaction (polymerasechainreaction).
SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sodium dodecylsulfate-polyacrylamide gel
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