BioTechniques最新文献_第3页

Who wants to live forever? Models shaping the future of aging research. 谁想长生不老？塑造老龄化研究未来的模型。

IF 2.2 4区工程技术

BioTechniques Pub Date : 2024-11-01 Epub Date: 2025-01-06 DOI: 10.1080/07366205.2024.2445467

Annie Coulson

引用次数: 0

Optimized for routine: highly sensitive fluorescent Telomeric Repeat Amplification Protocol (f-TRAP). 常规优化：高灵敏度荧光端粒重复扩增协议（f-TRAP）。

IF 2.2 4区工程技术

BioTechniques Pub Date : 2024-10-01 Epub Date: 2024-12-15 DOI: 10.1080/07366205.2024.2432805

Silke Fähnrich, Anne Wedemann, Laura Steenpass, Wilhelm Gerhard Dirks

{"title":"Optimized for routine: highly sensitive fluorescent Telomeric Repeat Amplification Protocol (f-TRAP).","authors":"Silke Fähnrich, Anne Wedemann, Laura Steenpass, Wilhelm Gerhard Dirks","doi":"10.1080/07366205.2024.2432805","DOIUrl":"10.1080/07366205.2024.2432805","url":null,"abstract":"The strict suppression of telomerase activity (TA) in terminally differentiated human cells causes a shortening of the chromosome ends after each cell division. This tumor suppression surveillance mechanism is associated with a limited number of cell divisions known as Hayflick limit. Here we present an optimized protocol for measuring TA that combines a fluorescently labeled bait primer and polymerase chain reaction (PCR) amplification with analytical capillary electrophoresis (CE) to achieve a detection limit of one telomerase-positive cell per ten thousand negative cells. In research laboratories today, a broad panel of TRAP assay protocols enables the assessment of the immortality of newly generated cell lines or the unambiguous evaluation of the reprogramming of induced pluripotent stem cells (iPSCs). The present f-TRAP protocol, optimized for routine use, enables fast ad hoc application for single measurements up to a high throughput of mass samples using a triplicate approach of different lysate concentrations. Final CE analysis facilitates standardized data processing and storage for documentation of results and could make f-TRAP a useful assay in research and clinical oncology.","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"517-522"},"PeriodicalIF":2.2,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142827093","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

Importance of a reference ladder in analytical testing of CRISPR HDR donor ssDNA templates. 参考阶梯在CRISPR HDR供体ssDNA模板分析测试中的重要性。

IF 2.2 4区工程技术

BioTechniques Pub Date : 2024-10-01 Epub Date: 2024-12-04 DOI: 10.1080/07366205.2024.2429308

Zhi-Xiang Lu, Anis H Khimani, Yanhong Tong

引用次数: 0

A single-cell 3D dynamic volume control system for chondrocytes. 软骨细胞单细胞三维动态体积控制系统

IF 2.2 4区工程技术

BioTechniques Pub Date : 2024-10-01 Epub Date: 2024-10-15 DOI: 10.1080/07366205.2024.2412414

Qiang Zhang, Yiyao Wang, Yanjun Zhang, Xiaochun Wei, Weiyi Chen, Quanyou Zhang

{"title":"A single-cell 3D dynamic volume control system for chondrocytes.","authors":"Qiang Zhang, Yiyao Wang, Yanjun Zhang, Xiaochun Wei, Weiyi Chen, Quanyou Zhang","doi":"10.1080/07366205.2024.2412414","DOIUrl":"10.1080/07366205.2024.2412414","url":null,"abstract":"In articular cartilage, zone-specific cellular morphology is a typical characteristic of cartilage tissue, which is related with chondrocyte function, inflammation and osteoarthritis (OA). Chondrocyte hypertrophic phenotype is a criticle physiological process which indicates a hallmark of chondrocyte terminal differentiation and bone formation. Thus, developing a in vitro cell culture system for dynamic regulation of single chondrocyte volume at a three-dimensional (3D) level is particularly necessary for understanding how physical cues of matrix microenvironment regulate chondrocyte fate and the degeneration of articular cartilage. Here, based on the soft lithography techniques, we have constructed well-defined single-cell 3D dynamic volume control system to recapitulate the physiological matrix microenvironment of single chondrocyte niche. The results of finite element analysis indicated that the stress and strain distribution in the cell culture region is homogeneous during the stretching process. Additionally, 3D dynamic volume expansion and compression of single cells in physiological or hyperphysiological can be realized in this cell culture system. Our device for single-cell 3D dynamic culture provides a microphysiological culture system for chondrocytes to explore the mechanisms of cartilage hypertrophy, as well as develops a new paradigm for functional cartilage tissue engineering and regenerative medicine.","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"495-504"},"PeriodicalIF":2.2,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142457178","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

Development and validation of a portable device for lab-free versatile nucleic acid extraction. 一种便携式无实验室多功能核酸提取装置的开发和验证。

IF 2.2 4区工程技术

BioTechniques Pub Date : 2024-10-01 Epub Date: 2024-12-02 DOI: 10.1080/07366205.2024.2427544

Anthony J Politza, Tianyi Liu, Aneesh Kshirsagar, Ming Dong, Md Ahasan Ahamed, Weihua Guan

{"title":"Development and validation of a portable device for lab-free versatile nucleic acid extraction.","authors":"Anthony J Politza, Tianyi Liu, Aneesh Kshirsagar, Ming Dong, Md Ahasan Ahamed, Weihua Guan","doi":"10.1080/07366205.2024.2427544","DOIUrl":"10.1080/07366205.2024.2427544","url":null,"abstract":"Nucleic acid testing (NAT) has revolutionized diagnostics by providing precise, rapid, and scalable detection methods for diverse biological samples. These recent advancements satisfy the increasing demand for on-site diagnostics, yet sample preparation remains a significant bottleneck for achieving highly sensitive diagnostic assays. There is an unmet need for compatible, efficient, and lab-free sample preparation for point-of-care NAT. To address this, we developed a portable, lab-free, and battery-powered device for extracting nucleic acids. We explored using low centrifugal forces with existing commercial chemistry, demonstrating excellent performance. We designed and tested a battery-powered device to enable lab-free extractions, and verified reagents stored out to 6 months, suggesting exceptional deployment capabilities. We evaluated our device, comparing our results against those from a benchtop centrifuge across three types of samples: HIV RNA in buffer, HIV RNA in plasma, and SARS-CoV-2 RNA in saliva. The portable device demonstrated excellent agreement with the benchtop centrifuge, indicating high reliability. By providing an effective on-site sample preparation solution, the widespread adoption of low centrifugal extractions will improve the sensitivity and reliability of NAT and will positively impact other point-of-care technologies such as next generation sequencing (NGS), biomarker detection, and environmental monitoring.","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"505-515"},"PeriodicalIF":2.2,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765762","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

CRISPR in 3D: Innovations in Disease Modelling and Personalized Medicine. 三维 CRISPR：疾病建模和个性化医疗的创新。

IF 2.2 4区工程技术

BioTechniques Pub Date : 2024-10-01 Epub Date: 2024-11-28 DOI: 10.1080/07366205.2024.2418748

Louis J Gautier

引用次数: 0

Comparison of commercially available DNA and RNA extraction kits for wildlife feces collected from the environment. 针对从环境中收集的野生动物粪便的市售 DNA 和 RNA 提取试剂盒比较。

IF 2.7 4区工程技术

BioTechniques Pub Date : 2024-09-13 DOI: 10.1080/07366205.2024.2397284

James D Feller,Leah Colton

引用次数: 0

A HABA dye-based colorimetric assay to detect unoccupied biotin binding sites in an avidin-containing fusion protein. 基于 HABA 染料的比色测定法，用于检测含阿维丁融合蛋白中未被占用的生物素结合位点。