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Correction. 修正。
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-09-18 DOI: 10.1080/07366205.2025.2563421
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引用次数: 0
Taking root: the techniques growing genetically engineered plants. 生根发芽：种植转基因植物的技术。
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-09-17 DOI: 10.1080/07366205.2025.2560832
Beatrice Bowlby
{"title":"Taking root: the techniques growing genetically engineered plants.","authors":"Beatrice Bowlby","doi":"10.1080/07366205.2025.2560832","DOIUrl":"https://doi.org/10.1080/07366205.2025.2560832","url":null,"abstract":"","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"1-5"},"PeriodicalIF":2.5,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145079598","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
Stabilization of human RNA and DNA in stool samples at room temperature. 室温下粪便样本中人类RNA和DNA的稳定。
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-08-18 DOI: 10.1080/07366205.2025.2546762
Regina Preywisch, Roger Løvlie, Moritz Eidens
{"title":"Stabilization of human RNA and DNA in stool samples at room temperature.","authors":"Regina Preywisch, Roger Løvlie, Moritz Eidens","doi":"10.1080/07366205.2025.2546762","DOIUrl":"https://doi.org/10.1080/07366205.2025.2546762","url":null,"abstract":"<p><p>This study examines the stability of human mRNA and DNA in stool samples for noninvasive gastrointestinal disease detection. While stool samples are valuable for diagnosing conditions like inflammatory disorders and colorectal cancer, mRNA instability poses significant challenges, risking false-negative results. To investigate this, 97 stool samples were treated with a specialized stabilizing solution and stored at room temperature, with analyses conducted on Day 1 and Day 15. The research aimed to improve storage protocols for enhanced reliability in mRNA diagnostics, aiding in personalized medicine and biomarker discovery. Results showed variability in total nucleic acid yields, increasing from Day 1 (mean 112 ng/µL) to Day 15 (mean 165 ng/µL), highlighting the benefits of improved homogenization and bacterial lysis. Human DNA remained stable over the 14-day period. For RNA stability, three mRNA markers were analyzed: Carcinoembryonic Antigen (CEACAM5), Prostaglandin-Endoperoxide Synthase 2 (PTGS2) and cortactin (CTTN). Both CEACAM5 (p=0.064) and PTGS2 (p=0.79) maintained stability, while CTTN showed a statistically significant but only modest reduction in expression (p < 0.0001). Overall, the stabilization buffer proved effectiveness in preserving nucleic acids and provided insights into mRNA marker stability over time.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"1-9"},"PeriodicalIF":2.5,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144871263","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
Nondestructive DNA extraction from specimens and bulk samples preserved in DESS solution for DNA barcoding. 从保存在DESS溶液中的标本和大样本中进行无损DNA提取，用于DNA条形码。
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-07-01 Epub Date: 2025-08-07 DOI: 10.1080/07366205.2025.2542023
Eri Ogiso-Tanaka, Minako Abe Ito, Daisuke Shimada
{"title":"Nondestructive DNA extraction from specimens and bulk samples preserved in DESS solution for DNA barcoding.","authors":"Eri Ogiso-Tanaka, Minako Abe Ito, Daisuke Shimada","doi":"10.1080/07366205.2025.2542023","DOIUrl":"10.1080/07366205.2025.2542023","url":null,"abstract":"<p><p>DNA barcoding of small organisms often requires significant damage or destroy specimens. To address this, the development of nondestructive DNA extraction methods that maintain specimen morphology is crucial. Here, we present a protocol using the supernatant of DESS preservation solution (20% DMSO, 250 mM EDTA, saturated NaCl), which conserve both the morphological characteristics and DNA of biological samples long-term. This method successfully conducted nondestructive barcoding of nematodes preserved in DESS and stored 10 years at room temperature. The protocol also applies to bulk environmental samples, where sediment and seagrass collected in the field are immediately preserved in DESS. This enables the subsequent isolation and individual nondestructive barcoding of meiofauna and diatoms from the preserved environmental samples in the laboratory.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"297-311"},"PeriodicalIF":2.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144793392","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 E. coli two-hybrid system to investigate human protein-protein interactions. 大肠杆菌双杂交系统研究人类蛋白质-蛋白质相互作用。
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-07-01 Epub Date: 2025-08-20 DOI: 10.1080/07366205.2025.2542028
Rebecca M Richardson, Sarah M Ho, Lane Tong, Morgan G Daniels, Steven M Pascal
{"title":"An <i>E. coli</i> two-hybrid system to investigate human protein-protein interactions.","authors":"Rebecca M Richardson, Sarah M Ho, Lane Tong, Morgan G Daniels, Steven M Pascal","doi":"10.1080/07366205.2025.2542028","DOIUrl":"10.1080/07366205.2025.2542028","url":null,"abstract":"<p><p>The LexA-<i>E. coli</i> two-hybrid (LexA-E2H) system was initially developed to study interactions between microbial proteins in an <i>Escherichia coli</i> (<i>E. coli)</i> environment. We here demonstrate its utility for studying mammalian protein interactions. Specifically, this study uses LexA-E2H to provide the first direct and quantitative validation of Glucose Regulated Protein 78 (GRP78) binding to the cleaved-Prostate Apoptosis Response 4 (cl-Par-4) tumor suppressor. Furthermore, the results establish that this interaction does not require phosphorylation of either protein. MacConkey agar was used for initial detection of the interaction through colorimetric colony screening, distinguishing pale white-pink colonies (+ interaction) from red colonies (- interaction). This was followed by β-galactosidase assays for quantitative assessment. These results demonstrate the potential of the LexA-E2H system to advance human protein-protein interaction research. LexA-E2H is simple to implement, avoiding the need to culture eukaryotic cells, and bypassing interference from eukaryotic proteins. This system is ideal for laboratories with limited resources and complements conventional eukaryotic methods.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"283-295"},"PeriodicalIF":2.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12430536/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144881989","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
Impact of ambient temperature exposure on miRNA stability in human plasma. 环境温度暴露对人血浆中miRNA稳定性的影响。
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-07-01 Epub Date: 2025-09-08 DOI: 10.1080/07366205.2025.2555657
Véronique Desgagné, Flore Lavoie, Imad Soukar, Marie-France Hivert, Luigi Bouchard, Perrie F O'Tierney-Ginn
{"title":"Impact of ambient temperature exposure on miRNA stability in human plasma.","authors":"Véronique Desgagné, Flore Lavoie, Imad Soukar, Marie-France Hivert, Luigi Bouchard, Perrie F O'Tierney-Ginn","doi":"10.1080/07366205.2025.2555657","DOIUrl":"10.1080/07366205.2025.2555657","url":null,"abstract":"<p><p>MicroRNAs (miRNAs) are considered more stable than mRNA, but the impact of progressive thawing of biological samples after freezing as may happen during shipping delays has not been quantified. To address this, we utilized digital PCR to estimate the absolute concentrations of select miRNAs following progressive thawing of human plasma and maintenance at ambient temperature. Specifically, we quantified let-7b-3p, miR-144-5p, miR-150-5p, miR-517a-3p, miR-524-5p, and miR-1283, which have varying abundance in plasma. We observed a trend indicating a decline in miRNA concentration as plasma samples were progressively thawed. Notably, miR-150-5p and miR-517a-3p were the least stable and were degraded by 32% and 52% respectively after 24 hours of ambient temperature storage. We found that the variation in sensitivity to temperature was not due to the GC content of the miRNAs nor their initial abundance, suggesting that other factors, such as protein interactors and vesicles carrying these miRNAs, may impact sensitivity.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"271-282"},"PeriodicalIF":2.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145013838","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
How do you measure the success and impact of a core facility? 你如何衡量一个核心设施的成功和影响？
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-07-01 Epub Date: 2025-09-11 DOI: 10.1080/07366205.2025.2555118
Kara A Clissold, Michelle S Itano
{"title":"How do you measure the success and impact of a core facility?","authors":"Kara A Clissold, Michelle S Itano","doi":"10.1080/07366205.2025.2555118","DOIUrl":"10.1080/07366205.2025.2555118","url":null,"abstract":"<p><p>What makes a successful core facility? While many metrics are suggested and requested to evaluate the success and impact of a core, a comprehensive understanding of the priority of the institution and core and how each metric fits into the overall strategy for the core facility is critical before determining research value or success in specific instances. A description of some different metrics commonly used to evaluate cores is presented in addition to the variable impact or interpretation of the metric in the case of different core facility structures. For example, in the case of a research - focused institution or core facility, the number and type of publications contributed to may be highly valued. Contrastingly, publications may be of less evaluative value in the case of a teaching or discovery - prioritized institution. Overall, we suggest a balanced view of core evaluation that presents each indicator in the specific context of the institution and specific core and technology.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"263-270"},"PeriodicalIF":2.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145032626","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
Optimizing chromosome yield: a comparative analysis of harvesting, preparation and waste recovery methods. 优化染色体产量：收获、制备和废物回收方法的比较分析。
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-05-01 Epub Date: 2025-08-11 DOI: 10.1080/07366205.2025.2536438
Sarah L Berger, Rinyaporn Phengchat, Stanley W Botchway, Mohammed Yusuf
{"title":"Optimizing chromosome yield: a comparative analysis of harvesting, preparation and waste recovery methods.","authors":"Sarah L Berger, Rinyaporn Phengchat, Stanley W Botchway, Mohammed Yusuf","doi":"10.1080/07366205.2025.2536438","DOIUrl":"10.1080/07366205.2025.2536438","url":null,"abstract":"<p><p>In mitotic chromosome preparation, it is crucial to maximize chromosome yield for downstream cytogenetic analysis. Using HeLa cells as a model adherent cell, we assessed and compared the recovery of chromosomes from the entire process as well as the fraction of chromosomes that would generally become discarded in the standardly used trypsinization and mitotic-shake-off chromosome preparation methods. A higher chromosome yield for polyamine (PA) and methanol acetic acid (MAA) chromosomes was achieved using the mitotic-shake-off method compared to trypsinization. Moreover, mitotic arrest using colcemid or nocodazole gave similar PA and MAA chromosome yields in the commonly collected fractions. Interestingly, when comparing the fractions that would usually be discarded in the mitotic-shake-off, for colcemid-treated cells compared to nocodazole-treated cells, a greater number of PA chromosomes was recovered from the former. Our results show that chromosomes can be retrieved from the waste media. These recovered chromosomes display a suitable morphology in all chromosome preparations, suggesting that in conditions where high chromosome yields are required, utilizing the mitotic-shake-off method and recovering the generally discarded chromosome fraction together with the commonly used fraction would aid in maximizing chromosome yield.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"245-256"},"PeriodicalIF":2.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144820482","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
Parameter optimization of NanoJ-SRRF for live-cell microtubule imaging. NanoJ-SRRF活细胞微管成像参数优化。
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-05-01 Epub Date: 2025-07-22 DOI: 10.1080/07366205.2025.2534301
Sanhua Fang, Li Liu, Dan Yang, Shuangshuang Liu, Junli Xuan, Guifeng Xiao, Qianbing Zhao
{"title":"Parameter optimization of NanoJ-SRRF for live-cell microtubule imaging.","authors":"Sanhua Fang, Li Liu, Dan Yang, Shuangshuang Liu, Junli Xuan, Guifeng Xiao, Qianbing Zhao","doi":"10.1080/07366205.2025.2534301","DOIUrl":"10.1080/07366205.2025.2534301","url":null,"abstract":"<p><p>Super-Resolution Radial Fluctuation (SRRF) enables live-cell super-resolution imaging, but requires careful parameter selection. Here, we quantify the impact of NanoJ-SRRF parameters on microtubule imaging using FWHM and SQUIRREL-based error mapping. Ring radius proved most critical, with values >1.0 degrading resolution and fidelity. Radiality magnification and axes in ring had minimal impact. Advanced parameters revealed pitfalls: \"remove positivity constraint\" degraded resolution by 43%, while gradient weighting catastrophically reduced fidelity (RSP = 0.204 ± 0.116). Temporal Radiality Average (TRA) outperformed Temporal Radiality Auto-Correlations (TRAC), milimizing artifacts. This study establishes the first evidence-based guidelines for live-cell tubulin imaging: ring radius ≤1.0, TRA mode prioritization, and avoidance of gradient weighting. Integrating FWHM and SQUIRREL offers a robust opitimization framework for cytoskeletal dynamics.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"231-243"},"PeriodicalIF":2.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144688795","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 simple and universal quasi-modular cloning system using NEBuilder® HiFi DNA assembly kit. 一个简单和通用的准模块化克隆系统使用NEBuilder®HiFi DNA组装套件。
IF 2.5 4区 工程技术
BioTechniques Pub Date : 2025-05-01 Epub Date: 2025-08-17 DOI: 10.1080/07366205.2025.2542012
M Rey Toleco, Mark van der Giezen
{"title":"A simple and universal quasi-modular cloning system using NEBuilder<sup>®</sup> HiFi DNA assembly kit.","authors":"M Rey Toleco, Mark van der Giezen","doi":"10.1080/07366205.2025.2542012","DOIUrl":"10.1080/07366205.2025.2542012","url":null,"abstract":"<p><p>Modular cloning has transformed synthetic biology by enabling rapid assembly of standardized DNA parts, but its effectiveness is often limited to well-characterised model organisms. Here, we expand the utility of the NEBuilder<sup>®</sup> HiFi DNA Assembly Kit (New England Biolabs) into a quasi-modular system that uses single-stranded oligonucleotide bridges to assemble reusable DNA fragments without the need for restriction enzymes. Using the anaerobic gut eukaryote <i>Blastocystis</i> ST7-B as a test case, we constructed and reassembled vectors with diverse promoter-terminator combinations. These results underscore the versatility of the method and its potential to accelerate genetic tool development in non-model biological systems.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"257-262"},"PeriodicalIF":2.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144862098","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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