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Fully in vitro iterative construction of a 24 kb-long artificial DNA sequence to store digital information. 完全体外迭代构建 24 kb 长的人工 DNA 序列，用于存储数字信息。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-04-04 DOI: 10.2144/btn-2023-0109
Julien Leblanc, Olivier Boulle, Emeline Roux, Jacques Nicolas, Dominique Lavenier, Yann Audic
{"title":"Fully in vitro iterative construction of a 24 kb-long artificial DNA sequence to store digital information.","authors":"Julien Leblanc, Olivier Boulle, Emeline Roux, Jacques Nicolas, Dominique Lavenier, Yann Audic","doi":"10.2144/btn-2023-0109","DOIUrl":"https://doi.org/10.2144/btn-2023-0109","url":null,"abstract":"In the absence of a DNA template, the <i>ab initio</i> production of long double-stranded DNA molecules of predefined sequences is particularly challenging. The DNA synthesis step remains a bottleneck for many applications such as functional assessment of ancestral genes, analysis of alternative splicing or DNA-based data storage. In this report we propose a fully <i>in vitro</i> protocol to generate very long double-stranded DNA molecules starting from commercially available short DNA blocks in less than 3 days using Golden Gate assembly. This innovative application allowed us to streamline the process to produce a 24 kb-long DNA molecule storing part of <i>the</i> <i>Declaration of the Rights of Man and of the Citizen</i> of 1789 . The DNA molecule produced can be readily cloned into a suitable host/vector system for amplification and selection.","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"37 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140571062","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
Considerations for developing mitochondrial transplantation techniques for individualized medicine. 开发用于个体化医疗的线粒体移植技术的考虑因素。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-04-01 Epub Date: 2024-02-29 DOI: 10.2144/btn-2023-0072
Kit Neikirk, Dominique C Stephens, Heather K Beasley, Andrea G Marshall, Jennifer A Gaddy, Steven M Damo, Antentor O Hinton
{"title":"Considerations for developing mitochondrial transplantation techniques for individualized medicine.","authors":"Kit Neikirk, Dominique C Stephens, Heather K Beasley, Andrea G Marshall, Jennifer A Gaddy, Steven M Damo, Antentor O Hinton","doi":"10.2144/btn-2023-0072","DOIUrl":"10.2144/btn-2023-0072","url":null,"abstract":"<p><p>Tweetable abstract Mitochondrial transplantation has been used to treat various diseases associated with mitochondrial dysfunction. Here, we highlight the considerations in quality control mechanisms that should be considered in the context of mitochondrial transplantation.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"125-134"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139989248","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
An industry perspective approach and control strategy for implementation of ready-to-use cells in bioassays: survey outcome and recommendations. 在生物测定中使用即用型细胞的行业视角方法和控制策略：调查结果和建议。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-04-01 Epub Date: 2024-02-09 DOI: 10.2144/btn-2023-0086
John R White, Sammina Ahmed, Jan Amstrup, Petra Bennington, Zhijie Jey Cheng, Peter Day, Keneshia K Haenssen, Adi Kozminsky-Atias, Eva Martinez, Esther Hq Ong, Isam Qahwash, Elaine Se Stokes, Jenny Wang
{"title":"An industry perspective approach and control strategy for implementation of ready-to-use cells in bioassays: survey outcome and recommendations.","authors":"John R White, Sammina Ahmed, Jan Amstrup, Petra Bennington, Zhijie Jey Cheng, Peter Day, Keneshia K Haenssen, Adi Kozminsky-Atias, Eva Martinez, Esther Hq Ong, Isam Qahwash, Elaine Se Stokes, Jenny Wang","doi":"10.2144/btn-2023-0086","DOIUrl":"10.2144/btn-2023-0086","url":null,"abstract":"<p><p>The BioPhorum Development Group is an industry collaboration enabling the sharing of common practices for the development of biopharmaceuticals. Bioassays are an important part of an analytical control system. Utilization of ready-to-use cells can increase operational flexibility and improve efficiency by providing frozen cell banks uniform stock while removing challenges associated with maintaining cultured cells. The BioPhorum Development Group-Bioassay workstream conducted an intercompany benchmarking survey and group discussions around the use of ready-to-use cells for bioassays. The results of the collaboration provide alignment on nomenclature, production, qualification and implementation of ready-to-use cells to support the assay life cycle.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"135-144"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139705986","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
High-throughput screening for a SARS-CoV-2 frameshifting inhibitor using a cell-free protein synthesis system. 利用无细胞蛋白质合成系统高通量筛选 SARS-CoV-2 换帧抑制剂。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-04-01 Epub Date: 2024-01-31 DOI: 10.2144/btn-2023-0102
Kodai Machida, Rin Tanaka, Seraya Miki, Shotaro Noseda, Mayumi Yuasa-Sunagawa, Hiroaki Imataka
{"title":"High-throughput screening for a SARS-CoV-2 frameshifting inhibitor using a cell-free protein synthesis system.","authors":"Kodai Machida, Rin Tanaka, Seraya Miki, Shotaro Noseda, Mayumi Yuasa-Sunagawa, Hiroaki Imataka","doi":"10.2144/btn-2023-0102","DOIUrl":"10.2144/btn-2023-0102","url":null,"abstract":"<p><p>Programmed-1 ribosomal frameshifting (-1 PRF) is a translational mechanism adopted by some viruses, including SARS-CoV-2. To find a compound that can inhibit -1 PRF in SARS-CoV-2, we set up a high-throughput screening system using a HeLa cell extract-derived cell-free protein synthesis (CFPS) system. A total of 32,000 compounds were individually incubated with the CFPS system programmed with a -1 PRF-EGFP template. Several compounds were observed to decrease the -1 PRF-driven fluorescence, and one of them had some suppressive effect on -1 PRF of a SARS-CoV-2 genome sequence in transfected cells. Thus the CFPS system can be used as a tool for a high-throughput screening of chemicals.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"161-168"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139641560","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
Multiparameter screen optimizes immunoprecipitation. 多参数筛选可优化免疫沉淀。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-04-01 Epub Date: 2024-02-29 DOI: 10.2144/btn-2023-0051
Shaoshuai Xie, Leila Saba, Hua Jiang, Omar R Bringas, Mehrnoosh Oghbaie, Luciano Di Stefano, Vadim Sherman, John LaCava
{"title":"Multiparameter screen optimizes immunoprecipitation.","authors":"Shaoshuai Xie, Leila Saba, Hua Jiang, Omar R Bringas, Mehrnoosh Oghbaie, Luciano Di Stefano, Vadim Sherman, John LaCava","doi":"10.2144/btn-2023-0051","DOIUrl":"10.2144/btn-2023-0051","url":null,"abstract":"<p><p>Immunoprecipitation (IP) coupled with mass spectrometry effectively maps protein-protein interactions when genome-wide, affinity-tagged cell collections are used. Such studies have recorded significant portions of the compositions of physiological protein complexes, providing draft 'interactomes'; yet many constituents of protein complexes still remain uncharted. This gap exists partly because high-throughput approaches cannot optimize each IP. A key challenge for IP optimization is stabilizing <i>in vivo</i> interactions during the transfer from cells to test tubes; failure to do so leads to the loss of genuine interactions during the IP and subsequent failure to detect. Our high-content screening method explores the relationship between <i>in vitro</i> chemical conditions and IP outcomes, enabling rapid empirical optimization of conditions for capturing target macromolecular assemblies.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"145-152"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11091867/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139995522","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Extracting high-quality RNA from formaldehyde-fixed naturally aged neuromusculoskeletal tissues. 从甲醛固定的天然老化神经肌肉骨骼组织中提取高质量 RNA。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-04-01 Epub Date: 2024-02-09 DOI: 10.2144/btn-2023-0092
Seth D Thompson, Rajeswari Pichika, Richard L Lieber, Mitra Lavasani
{"title":"Extracting high-quality RNA from formaldehyde-fixed naturally aged neuromusculoskeletal tissues.","authors":"Seth D Thompson, Rajeswari Pichika, Richard L Lieber, Mitra Lavasani","doi":"10.2144/btn-2023-0092","DOIUrl":"10.2144/btn-2023-0092","url":null,"abstract":"<p><p>Modern approaches to discovering molecular mechanisms and validating treatments for age-related neuromusculoskeletal dysfunction typically rely on high-throughput transcriptome analysis. Previously harvested and fixed tissues offer an incredible reservoir of untapped molecular information. However, obtaining RNA from such formaldehyde-fixed neuromusculoskeletal tissues, especially fibrotic aged tissues, is technically challenging and often results in RNA degradation, chemical modification and yield reduction, prohibiting further analysis. Therefore, we developed a protocol to extract high-quality RNA from formaldehyde-fixed brain, cartilage, muscle and peripheral nerve isolated from naturally aged mice. Isolated RNA produced reliable gene expression data comparable to fresh and flash-frozen tissues and was sensitive enough to detect age-related changes, making our protocol valuable to researchers in the field of aging.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"153-160"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10988388/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139705987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Recording the brain in vivo: emerging technologies for the exploration of mental health conditions. 活体记录大脑：探索精神健康状况的新兴技术。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-04-01 Epub Date: 2024-03-14 DOI: 10.2144/btn-2024-0013
Tristan Free
{"title":"Recording the brain <i>in vivo</i>: emerging technologies for the exploration of mental health conditions.","authors":"Tristan Free","doi":"10.2144/btn-2024-0013","DOIUrl":"10.2144/btn-2024-0013","url":null,"abstract":"<p><p>Standfirst Mounting interest in mental health conditions over the last two decades has been coupled with the increasing sophistication of techniques to study the brain <i>in vivo</i>. [Formula: see text].</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"121-124"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140118685","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
An optimal genomic DNA extraction method for shoots of four Dendrocalamus species based on membership function analysis. 基于成员函数分析的四种 Dendrocalamus 树种嫩枝基因组 DNA 最佳提取方法。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-03-01 Epub Date: 2023-12-22 DOI: 10.2144/btn-2023-0087
Yingchun Ma, Bowen Li, Dejia Yang, Shuguang Wang, Lixia Yu, Hui Zhan, Juan Li
{"title":"An optimal genomic DNA extraction method for shoots of four <i>Dendrocalamus</i> species based on membership function analysis.","authors":"Yingchun Ma, Bowen Li, Dejia Yang, Shuguang Wang, Lixia Yu, Hui Zhan, Juan Li","doi":"10.2144/btn-2023-0087","DOIUrl":"10.2144/btn-2023-0087","url":null,"abstract":"<p><p>High-quality genomic DNA extraction is fundamental for the study of gene cloning and expression in plants. Therefore, this study evaluated several methods for extracting genomic DNA from shoots of four <i>Dendrocalamus</i> species to determine the optimal technique. Genomic DNA was extracted using three different methods: a commercial DNA extraction kit method, a modified cetyltrimethylammonium bromide method and a sodium dodecyl sulfate method. A membership function analysis was employed to compare these methods. The results demonstrated that the commercial DNA extraction kit method was the most effective and comprehensive approach for extracting genomic DNA from shoots of four <i>Dendrocalamus</i> species. Furthermore, this study provided valuable insights into optimizing techniques for extracting genomic DNA in other bamboo species.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"94-103"},"PeriodicalIF":2.7,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138828137","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
Corrigendum. 更正。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-03-01 Epub Date: 2024-02-28 DOI: 10.2144/btn-2020-0153c1
{"title":"Corrigendum.","authors":"","doi":"10.2144/btn-2020-0153c1","DOIUrl":"10.2144/btn-2020-0153c1","url":null,"abstract":"","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"119"},"PeriodicalIF":2.7,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139982268","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
Corrigendum. 更正。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2024-03-01 Epub Date: 2024-02-28 DOI: 10.2144/btn-2022-0032c1
{"title":"Corrigendum.","authors":"","doi":"10.2144/btn-2022-0032c1","DOIUrl":"10.2144/btn-2022-0032c1","url":null,"abstract":"","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"120"},"PeriodicalIF":2.7,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139982269","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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