GB/T 34776-2017 in English

VALID

T4 DNA ligase-Detection methods of activity and impurities of enzyme

Issued on:2017-11-01
Implemented on:2018-05-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $160.00
$156.00

Standard No:	GB/T 34776-2017
Document status:	VALID
Title in English:	T4 DNA ligase-Detection methods of activity and impurities of enzyme
Title in Chinese:	T4 DNA连接酶酶活及杂质检测方法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2017-11-01
Implemented on:	2018-05-01
ICS Classification:	07.080-Biology. Botany. Zoology
Chinese Classification:	A40-Basic Subject in general
Professional Classification:	GB-National Standard
Related Keywords:	enzyme preparations t4 dna ligase activity detection method definition enzyme activity t4 dna ligase-detection methods dna ligase reaction buffer applicationt4 dna ligase
Related Topics:	enzyme dna Enzyme activity enzyme dna Impurity detection Impurity detection mass spectrometer dna+ Mass spectrometry + enzyme + dna testing provinces Collagenase activity Enzyme Impurity method GB/T 34776-2017 Rapid detection of chitinase activity Impurities and Secondary mass spectrometry enzyme Enzyme Mass Spectrometry Impurity detection. mass spectrometer Impurity standardization method Bromine impurity detection Quality inspection method Enzyme activity dnazyme Enzyme liquid Enzyme analysis and detection method Impurity standardization method collagenase enzyme enzyme pfu + enzyme collagenase activity detection of single stranded dna how to detect dna detection of tick dna Enzyme Activity Test Method Liquid phase method impurities fetal dna test live enzyme ssdna+detection+method dna/ Enzyme coat Enzymes And miscellaneous Collagenase dnaase dna hydroxylase Enzyme grams dna test earliest absolute enzyme activity Component impurity detection quality testing enzymatic synthesis enzyme separation dna ligase T4 ligase Ligase dnazyme 1 Enzyme mass spectrometry dna hybridization instrument dnazyme use dna hybridization Enzyme method Impurity Determination Calculation Method Detection of impurity components When was the dna test available t4 enzyme Flow cytometry enzyme activity assay Detection of impurity components Detection of impurities single stranded dna detection How to use the enzyme DNA enzyme ssdnazyme dna qualitative method Enzyme activity ELISA Chemiluminescent Detection Enzyme activity Detection of e3 ligase activity detection of single stranded dna dna+tm value ts enzyme assay foreign dna testing How to detect impurities dna quality method Impurity standard t4 connection time detection of dna by ultraviolet method dna ligase single strand Enzyme contamination dna hybridization instrument traditional t4 dnazyme detection Enzyme activity detection Liveness detection method enzyme linkage dna ligase single strand dna connection e3 ligase dna detection analysis Enzyme Activity Test Method Impurity detection method Impurity detection method How long is the t4 connection How to Test Enzyme Activity t4 enzyme four degree connection and 16 degree dna detection method Abnormal enzyme activity Exoenzyme dnazyme dnazyme i dnazyme method biphasic enzymatic method enzyme separation t4 polymerase dna ladder test t1 enzyme holoenzyme trace dna detection dnazyme type 1 Enzyme Activity Determination and Analysis Method DNA enzyme detection method DNA enzyme site ssDNA detection Phosphatase detection method GBT34776 GB/T 34776-2017 E2 ligase Method for Determining Enzyme Activity Enzyme activity assay method Detect protoplast viability DNA ligase activity DNA hybridization conditions Microplate reader stability testing method RRT for impurity testing DNA ligase ligation strategies Splint R ligase ligation principle src activity detection method Enzyme encapsulation method PKC enzyme expression detection Continuous detection method to measure enzyme activity How to measure od value with microplate reader Enzyme testing ingredients Micro DNA detection taq Dry chemical enzymatic testing industry Commercial enzyme activity assay Preparation and method of antibody enzyme Filter paper enzyme activity Enzyme activity measurement method Water activity detection method Method for detecting impurities on rubber block surface Ground detection method Telomerase activity detection method Catalase enzyme activity detection method Tag ligase quality Enzyme activity measurement method Water activity detection method Liver lipase detection method Detection method of activation degree Methods to detect the activity of protoplasts Adenosine deaminase detection method Lysosomal function detection method Common methods for measuring enzyme activity Enzyme kinetic assay method What are the lysosome detection methods? Luciferase activity assay method to detect enzyme activity The simplest way to measure enzyme activity Water activity detection method Impurity Control Methods Enzyme potency determination method Lysosomal enzyme activity detection method Methods to detect enzymatic reactions Method for determination of manganese peroxidase enzyme activity Enzyme labeling method Multifunctional enzyme assay Ethanol organic impurity detection method Kinase activity detection method How to measure water activity Examples of methods for measuring enzyme activity What are the methods for detecting impurities? Adenosine deaminase detection method Common testing methods for impurities Collagenase digestion method Microplate reader concentration detection method Enzyme labeling methods Luciferase Assay Method Luciferase luminescence detection method using microplate reader What method is used to measure enzyme activity? Enzyme deactivation method Impurity detection method robustness Cephalosporin activity assay method Lysosomal acid lipase detection method Methods for measuring enzyme activity include Enzyme preparation method Lysosome detection method Muscle biopsy enzyme staining test method Detection methods for impurities in rice What are the detection methods for enzymes in milk? Enzyme titer determination method Quartz sand impurity detection method Cutinase enzyme activity assay method Microplate reader assay method Method for measuring enzyme activity using microplate reader Water activity detection method Microplate reader measurement method dual wavelength method Lysosomal digestion function detection method Enzyme liquid impurity removal method Genotoxic impurity detection methods Mitochondrial activity detection method Methods for detecting impurities in pharmaceutical liquids Commonly used wavelength microplate reader detection methods Enzyme activity measurement method Adenosine deaminase detection method Water activity detection method Gas impurity detection Luciferase detection method Protoplast activity detection method telomerase activity Detection method of bacterial laccase What are the methods for measuring enzyme activity? Enzyme evolution method Methods for measuring polypeptide activity Enzyme activity Methods for measuring enzyme activity Rubidium iodide impurity detection Immobilized enzyme activity detection method Enzyme activity screening method Enzyme liquid extraction and determination methods Plasminogen detection method Cell telomerase activity detection method How to measure enzyme activity? Leaf photosynthetic enzyme activity assay Enzyme extraction method Single impurity test method What are the methods for measuring enzyme activity? What are the methods for enzyme activity determination? Methods for determining enzyme activity Methods for detecting enzyme activity Methods for detecting enzyme activity Methods for detecting enzyme activity

《GB/T 34776-2017T4 DNA连接酶酶活及杂质检测方法》由SWG11（全国工具酶标准化工作组）归口，主管部门为国家标准化管理委员会。

Introduction

In-depth interpretation of the National Standard of the People's Republic of China GB/T34776-2017

Standard dimensions	Comparison content
Scope of application	T4 DNA ligase for molecular biology research
Detection method	Enzyme activity determination, impurity detection (including nuclease, exonuclease, etc.)
Background of standard formulation	Respond to the needs of molecular biology research and application, and ensure the quality control of enzyme preparations

T4 DNA ligase activity detection method

Definition: T4 DNA ligase is an enzyme encoded by T4 bacteriophage gene 30, which relies on ATP to provide energy and catalyzes the formation of phosphodiester bonds between the 5' phosphate end and the 3'-hydroxyl end of double-stranded DNA or RNA.

Activity unit determination: In 20μL 1×T4 DNA ligase reaction buffer, the amount of enzyme required to connect 50% of the HindⅢ digested input DNA fragments (5' end concentration is 0.12μmol/L) within 30 minutes at 16℃ is 1 activity unit.

Reaction conditions	Specific requirements
Buffer composition	400 mmol/L Tris-Cl, 100 mmol/L MgCl2, 100 mmol/L DTT, 5 mmol/L ATP (pH7.8)
Reaction temperature	16℃
Reaction time	30 minutes

Impurity detection methods and technical points

Endonuclease detection

Steps: 1. Prepare ligation reaction system (Table 2); 2. Add substrate DNA (such as pBR322, pUC19, etc.); 3. React at the optimal temperature for 4 hours; 4. Check whether the bands are clear by electrophoresis to determine the endonuclease contamination.

Reagents	Sample volume
10×T4 DNA ligase reaction buffer	5μL
Substrate DNA	1μg
ddH2O	Make up to 50μL
Test enzyme	20 U

Background of standard formulation and analysis of technological evolution

Background: T4 DNA ligase plays an important role in molecular biology research and genetic engineering. Its quality control directly affects the reliability of experimental results.

Technological evolution: With the development of gene editing technology (such as CRISPR-Cas9), the demand for T4 DNA ligase has increased, which has promoted the continuous optimization and standardization of its detection methods.

Implementation recommendations

Reagent and instrument preparation: Ensure that all reagents and instruments meet the standard requirements (such as GB/T6682, YY/T0087, etc.).
Experimental operation specifications: Avoid contamination during the operation and strictly follow the temperature, time and other conditions.
Data recording and analysis: Record the electrophoresis results in detail, and evaluate the enzyme activity in combination with the calculation method in the standard appendix.
Quality control: Calibrate the instrument regularly to ensure the accuracy of the experimental results.

More samples: GB 18306-2015 JJG 21-2008 DL/T 5154-2012

Sample only — not a preview of GB/T 34776-2017

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