Yin Cheng Wong , Allan Wee Ren Ng , Andrew Osahor , Kumaran Narayanan
{"title":"Customizable BAC-based DNA markers for pulsed-field gel electrophoresis","authors":"Yin Cheng Wong , Allan Wee Ren Ng , Andrew Osahor , Kumaran Narayanan","doi":"10.1016/j.ab.2024.115596","DOIUrl":null,"url":null,"abstract":"<div><p>DNA markers are used as a size reference and sample loading control during gel electrophoresis. Most markers are designed for conventional gel electrophoresis to separate DNA smaller than 20 kb. For larger molecules, pulsed-field gel electrophoresis (PFGE) marker is required. Limited PFGE markers are available because large DNA are prone to nicking and degradation, causing smeary bands. Here, we developed a robust marker based on bacterial artificial chromosomes (BACs) with bands up to 184 kb. This marker could consistently confer intense and distinct bands for accurate gel analysis in molecular biology studies, laboratory validations or clinical diagnosis.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"693 ","pages":"Article 115596"},"PeriodicalIF":2.6000,"publicationDate":"2024-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0003269724001404/pdfft?md5=25b2edb62f29c593808d21043b930bf5&pid=1-s2.0-S0003269724001404-main.pdf","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical biochemistry","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0003269724001404","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
DNA markers are used as a size reference and sample loading control during gel electrophoresis. Most markers are designed for conventional gel electrophoresis to separate DNA smaller than 20 kb. For larger molecules, pulsed-field gel electrophoresis (PFGE) marker is required. Limited PFGE markers are available because large DNA are prone to nicking and degradation, causing smeary bands. Here, we developed a robust marker based on bacterial artificial chromosomes (BACs) with bands up to 184 kb. This marker could consistently confer intense and distinct bands for accurate gel analysis in molecular biology studies, laboratory validations or clinical diagnosis.
DNA 标记在凝胶电泳过程中用作大小参考和样品装载对照。大多数标记物设计用于传统凝胶电泳，以分离小于 20 kb 的 DNA。对于较大的分子，则需要脉冲场凝胶电泳（PFGE）标记。由于大 DNA 容易被切割和降解，从而造成涂抹带，因此目前可用的脉冲场凝胶电泳标记有限。在此，我们开发了一种基于细菌人工染色体（BAC）的稳健标记，其条带可达 184 kb。该标记可持续产生强烈而明显的条带，从而在分子生物学研究、实验室验证或临床诊断中进行准确的凝胶分析。
期刊介绍：
The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field.
The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology.
The journal has been particularly active in:
-Analytical techniques for biological molecules-
Aptamer selection and utilization-
Biosensors-
Chromatography-
Cloning, sequencing and mutagenesis-
Electrochemical methods-
Electrophoresis-
Enzyme characterization methods-
Immunological approaches-
Mass spectrometry of proteins and nucleic acids-
Metabolomics-
Nano level techniques-
Optical spectroscopy in all its forms.
The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.
京公网安备 11010802042870号
京ICP备2023020795号-1
分享
求助内容：
应助结果提醒方式：
扫码关注我们
