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PlayBack cloning: simple, reversible, cost-effective cloning for the combinatorial assembly of complex expression constructs. 回放克隆：简单、可逆、经济高效的克隆，用于复杂表达构建体的组合组装。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-10-01 Epub Date: 2023-10-10 DOI: 10.2144/btn-2023-0042
Gregory Foran, Ryan D Hallam, Marvel Megaly, Anel Turgambayeva, Aleksandar Necakov
{"title":"PlayBack cloning: simple, reversible, cost-effective cloning for the combinatorial assembly of complex expression constructs.","authors":"Gregory Foran, Ryan D Hallam, Marvel Megaly, Anel Turgambayeva, Aleksandar Necakov","doi":"10.2144/btn-2023-0042","DOIUrl":"10.2144/btn-2023-0042","url":null,"abstract":"<p><p>With advancements in multicomponent molecular biological tools, the need for versatile, rapid and cost-effective cloning that enables successful combinatorial assembly of DNA plasmids of interest is becoming increasingly important. Unfortunately, current cloning platforms fall short regarding affordability, ease of combinatorial assembly and, above all, the ability to iteratively remove individual cassettes at will. Herein we construct, implement and make available a broad set of cloning vectors, called PlayBack vectors, that allow for the expression of several different constructs simultaneously under separate promoters. Overall, this system is substantially cheaper than other multicomponent cloning systems, has usability for a wide breadth of experimental paradigms and includes the novel feature of being able to selectively remove components of interest at will at any stage of the cloning platform.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"168-178"},"PeriodicalIF":2.7,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41181906","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Single-cell RNA sequencing of bronchoscopy specimens: development of a rapid minimal handling protocol. 支气管镜标本的单细胞RNA测序：快速最小处理方案的开发。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-10-01 Epub Date: 2023-10-10 DOI: 10.2144/btn-2023-0017
Firoozeh V Gerayeli, Stephen Milne, Chen Xi Yang, Xuan Li, Elizabeth Guinto, Julia Shun-Wei Yang, Chung Yan Cheung, Tara R Stach, Tawimas Shaipanich, Janice M Leung, Don D Sin
{"title":"Single-cell RNA sequencing of bronchoscopy specimens: development of a rapid minimal handling protocol.","authors":"Firoozeh V Gerayeli, Stephen Milne, Chen Xi Yang, Xuan Li, Elizabeth Guinto, Julia Shun-Wei Yang, Chung Yan Cheung, Tara R Stach, Tawimas Shaipanich, Janice M Leung, Don D Sin","doi":"10.2144/btn-2023-0017","DOIUrl":"10.2144/btn-2023-0017","url":null,"abstract":"<p><p>Single-cell RNA sequencing (scRNA-seq) is an important tool for understanding disease pathophysiology, including airway diseases. Currently, the majority of scRNA-seq studies in airway diseases have used invasive methods (airway biopsy, surgical resection), which carry inherent risks and thus present a major limitation to scRNA-seq investigation of airway pathobiology. Bronchial brushing, where the airway mucosa is sampled using a cytological brush, is a viable, less invasive method of obtaining airway cells for scRNA-seq. Here we describe the development of a rapid and minimal handling protocol for preparing single-cell suspensions from bronchial brush specimens for scRNA-seq. Our optimized protocol maximizes cell recovery and cell quality and facilitates large-scale profiling of the airway transcriptome at single-cell resolution.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"157-167"},"PeriodicalIF":2.7,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41181907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Quantitative analysis of lncRNA in formalin-fixed paraffin-embedded tissues of oral squamous cell carcinoma. 口腔鳞状细胞癌福尔马林固定石蜡包埋组织中lncRNA的定量分析。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-10-01 Epub Date: 2023-08-17 DOI: 10.2144/btn-2023-0033
Kiran Kumar, Kaveri Hallikeri, Ajaykumar Oli, Mallikarjun Goni, Apoorva Jain, Jagadeesha Poyya, Alagilavada S Shilpasree, Palaksha Kanive Javaregowda
{"title":"Quantitative analysis of lncRNA in formalin-fixed paraffin-embedded tissues of oral squamous cell carcinoma.","authors":"Kiran Kumar,&nbsp;Kaveri Hallikeri,&nbsp;Ajaykumar Oli,&nbsp;Mallikarjun Goni,&nbsp;Apoorva Jain,&nbsp;Jagadeesha Poyya,&nbsp;Alagilavada S Shilpasree,&nbsp;Palaksha Kanive Javaregowda","doi":"10.2144/btn-2023-0033","DOIUrl":"10.2144/btn-2023-0033","url":null,"abstract":"<p><p>The study evaluated expression profiles of few regulatory lncRNAs in oral squamous cell carcinoma and normal mucosa adjacent to oral cancer using paired fresh frozen and formalin-fixed paraffin-embedded (FFPE) tissues stored at a different duration of time (1-5 years) using real-time quantitative PCR. The quantity and quality of total RNA isolated from FFPE tissues was less compared with that of fresh frozen tissues, which resulted in a noncorrelation of quantification cycle values. Following normalization, the expression of lncRNAs in the paired tissues did not differ significantly. The differential expression of the lncRNAs in the study was consistent with The Cancer Genome Atlas head and neck squamous cell carcinoma database. The study findings demonstrate the possibility of performing accurate quantitative analysis of lncRNAs using short amplicons and standardized real-time quantitative PCR assays in oral squamous cell carcinoma FFPE samples.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"133-142"},"PeriodicalIF":2.7,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10005505","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Peter and the Beanstalk: tackling the giant questions of soybean nodulation. 彼得和豆茎：解决大豆结瘤的巨大问题。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-09-01 Epub Date: 2023-08-17 DOI: 10.2144/btn-2023-0060
Peter Gresshoff
{"title":"Peter and the Beanstalk: tackling the giant questions of soybean nodulation.","authors":"Peter Gresshoff","doi":"10.2144/btn-2023-0060","DOIUrl":"10.2144/btn-2023-0060","url":null,"abstract":"<p><p>The following is an edited interview, carried out by the Journal Development Editor of <i>BioTechniques</i>, Ashling Cannon, with Peter Gresshoff (University of Queensland, UQ; Brisbane, Australia). Peter is a plant developmental geneticist, using molecular and genetic tools to understand the complexities of gene networks during the control of nodule formation in legumes. He was the Director of the Center of Excellence for Integrative Legume Research and is now an Emeritus Professor at UQ.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"75 3","pages":"95-98"},"PeriodicalIF":2.7,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10667503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Education in microscopy: taking a closer look with Jennifer Waters. 显微镜教育：与詹妮弗·沃特斯一起仔细观察。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-09-01 Epub Date: 2023-08-29 DOI: 10.2144/btn-2023-0079
Jennifer Waters
{"title":"Education in microscopy: taking a closer look with Jennifer Waters.","authors":"Jennifer Waters","doi":"10.2144/btn-2023-0079","DOIUrl":"10.2144/btn-2023-0079","url":null,"abstract":"<p><p>Jennifer Waters (Nikon Imaging Center, Harvard Medical School, MA, USA) spoke to Ebony Torrington (Future Science Group, London, UK) about her career in microscopy and the importance of education. Jennifer is the Director of the Nikon Imaging Center (NIC) at Harvard Medical School and a Chan Zuckerberg Initiative (CZI) Imaging Scientist. She was trained in cell biology and microscopy by Ted Salmon at University of North Carolina (UNC) at Chapel Hill (NC, USA), where her research focused on mitosis and cell cycle regulation. Jennifer's main focus today, in addition to running the NIC, is developing light microscopy educational resources.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"75 3","pages":"91-94"},"PeriodicalIF":2.7,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10304060","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Special Focus Issue - Celebrating 40 years of publishing lab techniques: looking back to look foreword. 焦点特刊-庆祝实验室技术出版40周年：回顾前言。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-09-01 DOI: 10.2144/btn-2023-0081
Beatrice Bowlby, Tristan Free, Annie Coulson, Aisha Al-Janabi, Ashling Cannon, Ebony Torrington
{"title":"Special Focus Issue - Celebrating 40 years of publishing lab techniques: looking back to look foreword.","authors":"Beatrice Bowlby, Tristan Free, Annie Coulson, Aisha Al-Janabi, Ashling Cannon, Ebony Torrington","doi":"10.2144/btn-2023-0081","DOIUrl":"10.2144/btn-2023-0081","url":null,"abstract":"","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"75 3","pages":"75-77"},"PeriodicalIF":2.7,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10303020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A practical approach to genome assembly and annotation of Basidiomycota using the example of Armillaria. 以蜜环菌为例进行担子菌门基因组组装和注释的实用方法。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-09-01 Epub Date: 2023-09-08 DOI: 10.2144/btn-2023-0023
Deborah L Narh Mensah, Brenda D Wingfield, Martin Pa Coetzee
{"title":"A practical approach to genome assembly and annotation of Basidiomycota using the example of <i>Armillaria</i>.","authors":"Deborah L Narh Mensah,&nbsp;Brenda D Wingfield,&nbsp;Martin Pa Coetzee","doi":"10.2144/btn-2023-0023","DOIUrl":"10.2144/btn-2023-0023","url":null,"abstract":"<p><p>Technological advancements in genome sequencing, assembly and annotation platforms and algorithms that resulted in several genomic studies have created an opportunity to further our understanding of the biology of phytopathogens, including <i>Armillaria</i> species. Most <i>Armillaria</i> species are facultative necrotrophs that cause root- and stem-rot, usually on woody plants, significantly impacting agriculture and forestry worldwide. Genome sequencing, assembly and annotation in terms of samples used and methods applied in <i>Armillaria</i> genome projects are evaluated in this review. Infographic guidelines and a database of resources to facilitate future <i>Armillaria</i> genome projects were developed. Knowledge gained from genomic studies of <i>Armillaria</i> species is summarized and prospects for further research are provided. This guide can be applied to other diploid and dikaryotic fungal genomics.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"75 3","pages":"115-128"},"PeriodicalIF":2.7,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10297424","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Unraveling crime scenes strand by strand: the forensic odyssey of Bruce Budowle. 逐一揭开犯罪现场：布鲁斯·布道尔的法医之旅。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-09-01 Epub Date: 2023-08-25 DOI: 10.2144/btn-2023-0069
Bruce Budowle
{"title":"Unraveling crime scenes strand by strand: the forensic odyssey of Bruce Budowle.","authors":"Bruce Budowle","doi":"10.2144/btn-2023-0069","DOIUrl":"10.2144/btn-2023-0069","url":null,"abstract":"<p><p><b>Bruce Budowle speaks to Ashling Cannon, Journal Development Editor for <i>BioTechniques</i>, about advancements & challenges in forensic science.</b> Budowle completed his doctorate in genetics at Virginia Tech (VA, USA) formally known as Virginia Polytechnic Institute and State University. He then went on to complete a postdoctoral fellowship at the University of Alabama at Birmingham (AL, USA) to study genetic risk factors for acute lymphocytic leukemia, diabetes and melanoma. Budowle was early in his career and hadn't spent much time in forensics at this stage, but in 1982 an advert caught his eye for a job with the FBI to develop genetic marker systems to identify people who have left biological evidence at crime scenes. Budowle spent 26 years with the FBI and helped develop a plethora of genetic analysis methods. In 1985, it became a reality that DNA could be a signature for identifying people, and there were huge developments in DNA forensic analysis. In 2009, Budowle moved into academia and went to the University of North Texas Health Science Center (TX, USA), eventually becoming the Director of the Center for Human Identification, where he oversaw missing person and traditional crime cases, taught students and carried out fundamental and applied research. Budowle feels incredibly lucky to have had the resources, opportunities and academic infrastructure to learn and develop his knowledge. Budowle recently retired from academia and now spends his time building capacity for DNA forensics applications in Africa through the Department of Justice, with a well-established program known as the International Criminal Investigative Training Assistance Program (ICITAP) as well as with the non-government organization (NGO) DNAforAfrica.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"75 3","pages":"85-89"},"PeriodicalIF":2.7,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10303355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Western blotting (immunoblotting): history, theory, uses, protocol and problems. 蛋白质印迹（免疫印迹）：历史、理论、用途、方案和问题。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-09-01 Epub Date: 2023-03-27 DOI: 10.2144/btn-2022-0034
Rasheed Sule, Gabriela Rivera, Aldrin V Gomes
{"title":"Western blotting (immunoblotting): history, theory, uses, protocol and problems.","authors":"Rasheed Sule,&nbsp;Gabriela Rivera,&nbsp;Aldrin V Gomes","doi":"10.2144/btn-2022-0034","DOIUrl":"10.2144/btn-2022-0034","url":null,"abstract":"<p><p>Western blotting (immunoblotting) is a powerful and commonly used technique that is capable of detecting or semiquantifying an individual protein from complex mixtures of proteins extracted from cells or tissues. The history surrounding the origin of western blotting, the theory behind the western blotting technique, a comprehensive protocol and the uses of western blotting are presented. Lesser known and significant problems in the western blotting field and troubleshooting of common problems are highlighted and discussed. This work is a comprehensive primer and guide for new western blotting researchers and those interested in a better understanding of the technique or getting better results.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"75 3","pages":"99-114"},"PeriodicalIF":2.7,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10643815","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
I know what you're thinking; can neuroimaging truly reveal our innermost thoughts? 我知道你在想什么；神经影像学能真正揭示我们内心的想法吗？
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-09-01 Epub Date: 2023-08-25 DOI: 10.2144/btn-2023-0066
Jenny Straiton
{"title":"I know what you're thinking; can neuroimaging truly reveal our innermost thoughts?","authors":"Jenny Straiton","doi":"10.2144/btn-2023-0066","DOIUrl":"10.2144/btn-2023-0066","url":null,"abstract":"<p><p>[Formula: see text] Advances in neuroimaging, combined with developments in artificial intelligence software, have allowed researchers to noninvasively decode the brain and 'read the mind'.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"75 3","pages":"81-83"},"PeriodicalIF":2.7,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10307206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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