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GB/T 44829-2024: Assay method of Pf Ago endonuclease activity
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivered in | Standard Title (Description) | Status | PDF |
| GB/T 44829-2024 | English | 269 |
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Assay method of Pf Ago endonuclease activity
| Valid |
GB/T 44829-2024
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Standard similar to GB/T 44829-2024 GB/T 51033 GB/T 50082 Basic data | Standard ID | GB/T 44829-2024 (GB/T44829-2024) | | Description (Translated English) | Assay method of Pf Ago endonuclease activity | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | C27 | | Classification of International Standard | 07.080 | | Word Count Estimation | 13,169 | | Date of Issue | 2024-10-26 | | Date of Implementation | 2024-10-26 | | Issuing agency(ies) | State Administration for Market Regulation, China National Standardization Administration | GB/T 44829-2024: Assay method of Pf Ago endonuclease activity---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GBT44829-2024
ICS 07.080
CCSC27
National Standard of the People's Republic of China
PfAgo endonuclease activity detection method
Released on October 26, 2024
Implementation on October 26, 2024
State Administration for Market Regulation
The National Standardization Administration issued
Table of Contents
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 Test Step 2
7 Quality Control 7
Preface
This document is in accordance with the provisions of GB/T 1.1-2020 "Guidelines for standardization work Part 1.Structure and drafting rules for standardization documents"
Drafting.
Please note that some of the contents of this document may involve patents. The issuing organization of this document does not assume the responsibility for identifying patents.
This document was proposed and coordinated by the National Tool Enzyme Standardization Working Group (SAC/SWG11).
This document was drafted by. Jiaohong Biotechnology (Shanghai) Co., Ltd., Feipeng Biotechnology Co., Ltd., Nansheng (Xiamen) Analytical Testing Co., Ltd.
Co., Ltd., Xiahe (Shenzhen) Biotechnology Co., Ltd., Xiamen Yinxiang Group Co., Ltd., Shenzhen New Industry Biomedical Engineering Co., Ltd.
Company, Wuhan Xinhuayang Biotechnology Co., Ltd., Langfang Nordo Zhongke Medical Testing Laboratory Co., Ltd., Jiaohong Bioengineering (Shanghai) Co., Ltd.
Ltd., Xiahe (Hangzhou) Biotechnology Co., Ltd., Fujian Nansheng Technology Co., Ltd., Shanghai Jiaotong University, Fudan University, Shandong University, Suzhou
Zhou University, Xiamen University, Fujian Agriculture and Forestry University, and Xiamen Huaxia College.
The main drafters of this document are. Feng Yan, Li Zhonglei, Huang Facan, Zheng Dengzhong, Meng Yuan, Zhang Zhigang, Du Kai, Xu Li, Pang Tieliang, Zhao Yi, Chen Lishi,
Guo Yanwei, Liu Qian, Pan Wei, Ye Xingyu, Sun Peng, Zhong Jiang, Chen Xiulan, Fang Zhengwei, Yu Haiyue, Yu Xin, Deng Yongjiang, Zhu Li, Zhang Yongyou, Liu Bin,
Zhao Chao, Zheng Tianye, Huang Enming, Zhou Zhongai, Pan Paifeng, Huang Haiyan, Su Jingmei.
introduction
PfAgo endonuclease can cleave the DNA guide strand (greater than 15 nt) under the guidance of the 5' phosphorylated DNA guide strand.
The target DNA to be complemented, the cleavage site is generally at the 10th and 11th bases corresponding to the target DNA starting from the 5' end of the DNA guide strand
The cleavage function of PfAgo endonuclease is not limited by the target DNA and the adjacent base sequence of the cleavage site.
The activity of PfAgo endonuclease is relatively high and it also has some activity against target RNA substrates.
The establishment of national standards to promote their industrialization is of great significance for the production and use of this type of tool enzymes.
PfAgo endonuclease activity detection method
1 Scope
This document describes a method for detecting PfAgo endonuclease activity.
This document is applicable to the detection of PfAgo endonuclease activity.
2 Normative references
The contents of the following documents constitute the essential clauses of this document through normative references in this document.
For referenced documents without a date, only the version corresponding to that date applies to this document; for referenced documents without a date, the latest version (including all amendments) applies to
This document.
GB/T 191-2008 Pictorial symbols for packaging, storage and transportation
GB/T 6682 Specifications and test methods for water used in analytical laboratories
3 Terms and Definitions
The following terms and definitions apply to this document.
3.1
PfAgoendonuclease
Based on the complementary base pairing between the DNA guide strand (gDNA) sequence and the target DNA (tDNA) sequence
Yes, then the endonuclease that specifically recognizes and cuts the target DNA.
3.2
In a 25 μL reaction volume, at 95°C, the amount of enzyme required to cleave 0.1 nmol of target DNA per minute is one activity unit (U).
4 Principles
Based on the complementary base pairing between the DNA guide strand sequence and the target DNA sequence, PfAgo endonuclease can phosphorylate the
The target DNA complementary to the DNA guide strand is cut under the guidance of the DNA guide strand (greater than 15 nt). The cutting site often occurs at the end of the DNA guide strand.
The guide strand is complementary to the target DNA at the phosphodiester bond between bases 10 and 11 of the target DNA.
5 Reagents or materials
5.1 Water
Grade II water in accordance with GB/T 6682.
5.2 1mol/LTris-HCl (pH 8.0)
Weigh 12.114 g of Tris, add it to 80 mL of ultrapure water, mix thoroughly, adjust the pH to 8.0 ± 0.05 with 6 mol/L hydrochloric acid, and add
Add ultrapure water to a volume of 100.0 mL and filter with a 0.22 μm microporous filter membrane. Store at 2°C to 8°C. The shelf life is 24 months.
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