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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":null,"pages":null},"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":null,"pages":null},"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
HIV: the pursuit of an elusive vaccine. 艾滋病毒:寻找难以捉摸的疫苗。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-08-01 DOI: 10.2144/btn-2023-0059
Annie Coulson
{"title":"HIV: the pursuit of an elusive vaccine.","authors":"Annie Coulson","doi":"10.2144/btn-2023-0059","DOIUrl":"https://doi.org/10.2144/btn-2023-0059","url":null,"abstract":"<p><p>In 1984, when it was discovered that the HIV-1 virus caused AIDS, the US Health and Human Services Secretary, Margaret Heckler, declared that a vaccine would be available within 2 years. So why, 40 years on, are we still searching for an HIV vaccine? [Formula: see text].</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10047153","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
Releasing secrets bound to ancient remains with modern DNA extraction techniques: an interview with Elena Essel. 用现代 DNA 提取技术揭开古代遗骸的秘密：Elena Essel 的访谈。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-08-01 Epub Date: 2023-08-17 DOI: 10.2144/btn-2023-0067
Elena Essel
{"title":"Releasing secrets bound to ancient remains with modern DNA extraction techniques: an interview with Elena Essel.","authors":"Elena Essel","doi":"10.2144/btn-2023-0067","DOIUrl":"10.2144/btn-2023-0067","url":null,"abstract":"<p><p>Elena Essel (Msc) spoke to Ebony Torrington, Managing Editor of <i>BioTechniques</i>. Essel is a molecular biologist in Matthias Meyer's Advanced DNA Sequencing Techniques group at the Max Planck Institute for Evolutionary Anthropology in Leipzig (Germany). Essel studied biology at University of Erlangen-Nuremberg (Erlangen, Germany) for her bachelor's and in Martin-Luther-University Halle-Wittenberg (Halle an der Saale, Germany) for her master's. Essel worked in Meyer's group on DNA extraction of very degraded material for her master's thesis. Meyer is an expert in developing new cutting-edge methods for researching ancient DNA, with a focus on skeletal remains, and more recently on sediment remains. Essel now focusses on DNA sampling and extraction aspects of the pipeline at Meyer's lab for the ancient DNA workflow.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10048101","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
Selective expansion of target cells using the Enrich TROVO platform. 利用 Enrich TROVO 平台选择性扩增靶细胞。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-08-01 Epub Date: 2023-08-08 DOI: 10.2144/btn-2023-0038
Prem Shrestha, Stephen Rotatori, Kirby Madden-Hennessey, Christina Mohammed, Chi-Han Yang, Jordan Urbani, Joseph Pettinelli, Xueqi Liu, Qi Zhao
{"title":"Selective expansion of target cells using the Enrich TROVO platform.","authors":"Prem Shrestha, Stephen Rotatori, Kirby Madden-Hennessey, Christina Mohammed, Chi-Han Yang, Jordan Urbani, Joseph Pettinelli, Xueqi Liu, Qi Zhao","doi":"10.2144/btn-2023-0038","DOIUrl":"10.2144/btn-2023-0038","url":null,"abstract":"<p><p>Enriching target cell clones from diverse cell populations is vital for many life science applications. We have developed a novel method to rapidly and efficiently purify specific clonal cell populations from a larger, heterogeneous group using the Enrich TroVo system (Enrich Biosystems Inc., CT, USA). This system takes advantage of microfabrication and optical technologies by utilizing small hydrogel wells to separate desired cell populations and an innovative patching technique to selectively eliminate undesired cells. This method allows the isolation and growth of desired cells with minimal impact on their viability and proliferation. We successfully isolated and expanded clonal cell populations of desired cells using two model cells. Compared with fluorescence-activated cell sorting, Enrich TroVo system offers a promising alternative for isolating of sensitive, adherent cells, that is, patient-derived cells.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/79/3c/btn-75-56.PMC10476488.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10513046","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
A high-throughput pipeline for DNA/RNA/small RNA purification from tissue samples for sequencing. 从组织样品中纯化DNA/RNA/小RNA用于测序的高通量管道。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-08-01 DOI: 10.2144/btn-2023-0011
Jing Xu, Pawan K Pandoh, Richard D Corbett, Duane Smailus, Reanne Bowlby, Denise Brooks, Helen McDonald, Simon Haile, Sundeep Chahal, Steve Bilobram, Karen L Mungall, Andrew J Mungall, Robin Coope, Richard A Moore, Yongjun Zhao, Steven Jm Jones, Marco A Marra
{"title":"A high-throughput pipeline for DNA/RNA/small RNA purification from tissue samples for sequencing.","authors":"Jing Xu,&nbsp;Pawan K Pandoh,&nbsp;Richard D Corbett,&nbsp;Duane Smailus,&nbsp;Reanne Bowlby,&nbsp;Denise Brooks,&nbsp;Helen McDonald,&nbsp;Simon Haile,&nbsp;Sundeep Chahal,&nbsp;Steve Bilobram,&nbsp;Karen L Mungall,&nbsp;Andrew J Mungall,&nbsp;Robin Coope,&nbsp;Richard A Moore,&nbsp;Yongjun Zhao,&nbsp;Steven Jm Jones,&nbsp;Marco A Marra","doi":"10.2144/btn-2023-0011","DOIUrl":"https://doi.org/10.2144/btn-2023-0011","url":null,"abstract":"<p><p>High-throughput total nucleic acid (TNA) purification methods based on solid-phase reversible immobilization (SPRI) beads produce TNA suitable for both genomic and transcriptomic applications. Even so, small RNA species, including miRNA, bind weakly to SPRI beads under standard TNA purification conditions, necessitating a separate workflow using column-based methods that are difficult to automate. Here, an SPRI-based high-throughput TNA purification protocol that recovers DNA, RNA and small RNA, called GSC-modified RLT+ Aline bead-based protocol (GRAB-ALL), which incorporates modifications to enhance small RNA recovery is presented. GRAB-ALL was benchmarked against existing nucleic acid purification workflows and GRAB-ALL efficiently purifies TNA, including small RNA, for next-generation sequencing applications in a plate-based format suitable for automated high-throughput sample preparation.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10058280","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 novel use of a needle-free injection system for improved nucleic acid delivery and expression in vivo. 一种新型的无针注射系统，用于改善核酸在体内的传递和表达。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-08-01 DOI: 10.2144/btn-2023-0015
Nathan Liu, Hani Abd-Ul-Salam, Noémie Joannette-Lafrance, Jingjing Li, Karim Menassa, Monzur Murshed
{"title":"A novel use of a needle-free injection system for improved nucleic acid delivery and expression <i>in vivo</i>.","authors":"Nathan Liu,&nbsp;Hani Abd-Ul-Salam,&nbsp;Noémie Joannette-Lafrance,&nbsp;Jingjing Li,&nbsp;Karim Menassa,&nbsp;Monzur Murshed","doi":"10.2144/btn-2023-0015","DOIUrl":"https://doi.org/10.2144/btn-2023-0015","url":null,"abstract":"<p><p>Transfection, a nonviral method of nucleic acid delivery, often exhibits poor efficiency <i>in vivo</i>. The needle-based <i>in vivo</i> delivery of transfection reagents can be invasive. Here, we report a noninvasive protocol for <i>in vivo</i> gene delivery via the needle-free MED-JET H4 MULTIJET (MJH4M) device using both \"home-made\" glucose-based and commercial transfection reagents. The objective of this study was to compare the relative transfection efficiencies of the needle-free system to that of the needle-based delivery method. We observed a 15-fold increase in transfection efficiency using the needle-free MJH4M device when compared to the needle-based delivery method. The highest transfection efficiency was achieved using a 5% glucose solution as the delivery vehicle.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10057766","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
Chromatin immunoprecipitation and library preparation: a powerful tool to unravel the epigenome. 染色质免疫沉淀和文库制备:揭示表观基因组的有力工具。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-07-01 DOI: 10.2144/btn-2022-0118
Mukesh Pratap Yadav, Vikas Kundra
{"title":"Chromatin immunoprecipitation and library preparation: a powerful tool to unravel the epigenome.","authors":"Mukesh Pratap Yadav,&nbsp;Vikas Kundra","doi":"10.2144/btn-2022-0118","DOIUrl":"https://doi.org/10.2144/btn-2022-0118","url":null,"abstract":"","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9882904","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
The root microbiome: techniques for exploration and agricultural applications. 根系微生物组:探索和农业应用技术。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-07-01 DOI: 10.2144/btn-2023-0057
Ashling Cannon
{"title":"The root microbiome: techniques for exploration and agricultural applications.","authors":"Ashling Cannon","doi":"10.2144/btn-2023-0057","DOIUrl":"https://doi.org/10.2144/btn-2023-0057","url":null,"abstract":"<p><p>Standfirst With an ever-growing global population, the pursuit of sustainable agriculture has become paramount. What if the solution lies right beneath our feet? Enter the root microbiome, the hidden hero poised to revolutionize agriculture and bring us closer to a greener future. [Formula: see text].</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9883899","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
Overlap extension cloning of PCR products into a Gateway-compatible plasmid vector. PCR产物在gateway相容质粒载体上的重叠延伸克隆。
IF 2.7 4区 工程技术
BioTechniques Pub Date : 2023-07-01 DOI: 10.2144/btn-2023-0001
Omid Zare-Mehrjerdi, Gracie Trader, Viktor Kirik
{"title":"Overlap extension cloning of PCR products into a Gateway-compatible plasmid vector.","authors":"Omid Zare-Mehrjerdi,&nbsp;Gracie Trader,&nbsp;Viktor Kirik","doi":"10.2144/btn-2023-0001","DOIUrl":"https://doi.org/10.2144/btn-2023-0001","url":null,"abstract":"A PCR cloning method that combines a dual selection pGATE-1 plasmid vector and an improved overlap extension cloning was developed. This efficient and cost-effective method allows for the introduction of DNA fragments into the Gateway cloning pipeline. The cloning efficiency is facilitated by a dual selection that includes the ccdB gene and gentamicin resistance. For users of the Gateway cloning system, substantial cost saving comes from eliminating BP recombination and ligation reactions to introduce DNA fragments into pDONR or pENTR vectors. Beyond the Gateway technology, this recombination-based cloning system can be used to efficiently clone PCR amplicons by adding 24-base pair adaptor sequences that are utilized by bacterial homologous recombination mechanism.","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/61/ea/btn-75-363.PMC10388215.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9972567","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
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