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Transitioning from Triton X-100 to Tergitol 15-S-9: impacts on diagnostic assays using viral PCR sample solution. 从 Triton X-100 过渡到 Tergitol 15-S-9：对使用病毒 PCR 样品溶液进行诊断测定的影响。
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-05-05 DOI: 10.1080/07366205.2024.2342172
Azul Zorzoli, Alasdair MacLean, Scott Nicholson, Alison Daniels, Stephen Hughes, Susan Bennet-Slater, Christine Tait-Burkard, Noha El Sakka, Rory Gunson, Kate Templeton
{"title":"Transitioning from Triton X-100 to Tergitol 15-S-9: impacts on diagnostic assays using viral PCR sample solution.","authors":"Azul Zorzoli, Alasdair MacLean, Scott Nicholson, Alison Daniels, Stephen Hughes, Susan Bennet-Slater, Christine Tait-Burkard, Noha El Sakka, Rory Gunson, Kate Templeton","doi":"10.1080/07366205.2024.2342172","DOIUrl":"10.1080/07366205.2024.2342172","url":null,"abstract":"<p><p>In 2019, the European Union banned Triton X-100, a detergent widely used in laboratory diagnostics, including the Viral PCR Sample Solution (VPSS), and urged manufacturers to find environmentally sustainable alternatives. Tergitol 15-S-9 (VPSS2) has been proposed as an alternative surfactant. This multicenter study evaluated the effectiveness of VPSS2, a Tergitol-based viral solution, as a replacement for VPSS. Our results show the equivalent performance of VPSS2 to VPSS for nucleic acid extraction and viral stability over time at different temperatures. The new VPSS formulation was also tested against external quality assurance panels and clinical samples. The results of this work support adopting this modified viral PCR sample solution to replace Triton X-100-containing viral transport solutions.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"295-298"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140847473","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
Adipocyte ABCA1 expression analysis using flow cytometry. 使用流式细胞仪分析脂肪细胞 ABCA1 的表达。
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-07-17 DOI: 10.1080/07366205.2024.2376466
Sakshi Shukla, Ashutosh Bansal, Sandeep Aggarwal, Archna Singh
{"title":"Adipocyte ABCA1 expression analysis using flow cytometry.","authors":"Sakshi Shukla, Ashutosh Bansal, Sandeep Aggarwal, Archna Singh","doi":"10.1080/07366205.2024.2376466","DOIUrl":"10.1080/07366205.2024.2376466","url":null,"abstract":"<p><p>Adipocyte characterization and assessing membrane proteins using flow cytometry has been proven to be challenging as adipocytes are fragile, especially in subjects with high BMI. We overcame these challenges through a protocol optimizing tissue digestion time by reducing intermediate steps to minimize adipocyte friction and breakage. We avoided requirement for specialized instrument configuration and used a modified gating strategy to prevent inclusion of lipid droplets during analysis. Up to 90% of the cell population were available in the gating area. We checked the expression level of ABCA1, a membrane protein reaffirming adipocyte selection. In summary, this protocol requires lesser tissue sample improving feasibility and cost efficiency. Thus, our flow cytometry method is an improvement for studying adipocyte membrane characteristics.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"405-410"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141625897","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
Dual-mode detection of glucose based on pistol-like DNAzyme-mediated exonuclease-assisted signal cycle. 基于手枪式 DNA 酶介导的外切酶辅助信号循环的葡萄糖双模式检测。
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-08-05 DOI: 10.1080/07366205.2024.2381403
Huiying Lu, Xiaofeng Wang
{"title":"Dual-mode detection of glucose based on pistol-like DNAzyme-mediated exonuclease-assisted signal cycle.","authors":"Huiying Lu, Xiaofeng Wang","doi":"10.1080/07366205.2024.2381403","DOIUrl":"10.1080/07366205.2024.2381403","url":null,"abstract":"<p><p>Detecting glucose accurately and sensitively from clinical samples like tears and saliva is still difficult. We have created a sensor that can detect glucose with high sensitivity and accuracy by combining the use of glucose oxidase (GOx) to catalyze glucose, a pistol-like DNAzyme (PLDz) to transform the signal, gold nanoparticles (AuNPs) to enhance the optical properties and the exonuclease-III (Exo-III) to amplify the signal. As a result, the proposed method exhibits a low detection limit of 7.5 pM and a wide detection range covering seven orders of magnitude. The suggested dual-mode strategy provides a sensitive, precise and specific detection method for glucose. Another advantage is that the dual-mode technique significantly improves the precision and consistency of the measurements, demonstrating its immense potential for use in biomedical research and clinical diagnostics.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"415-423"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141888420","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 dynamic duo: cryo-EM teams up with machine learning to visualize biomolecules in motion. 动态二重奏：低温电子显微镜与机器学习相结合，实现生物分子运动的可视化。
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-06-12 DOI: 10.1080/07366205.2024.2355771
Beatrice Bowlby
{"title":"The dynamic duo: cryo-EM teams up with machine learning to visualize biomolecules in motion.","authors":"Beatrice Bowlby","doi":"10.1080/07366205.2024.2355771","DOIUrl":"10.1080/07366205.2024.2355771","url":null,"abstract":"<p><p>Cryo-EM has been a key technique in our understanding of biomolecular structures. Now, machine learning techniques are being used to put these structures in motion, revealing dynamic interactions and processes happening on a molecular and cellular level.[Formula: see text].</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"229-233"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141305338","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
Artificial intelligence: help or hindrance in solving the reproducibility crisis? 人工智能：帮助还是阻碍解决可重复性危机？
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-06-20 DOI: 10.1080/07366205.2024.2355776
Jennifer Straiton
{"title":"Artificial intelligence: help or hindrance in solving the reproducibility crisis?","authors":"Jennifer Straiton","doi":"10.1080/07366205.2024.2355776","DOIUrl":"10.1080/07366205.2024.2355776","url":null,"abstract":"<p><p>Science is in the midst of a reproducibility crisis and the integration of artificial intelligence into scientific research has only compounded the problem; yet could the technology hold the solution to its own problems?[Formula: see text].</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"291-294"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141426260","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
Simple and sensitive detection of Pseudomonas aeruginosa in neonatal infection based on a both-end blocked peroxidase-mimicking DNAzyme. 基于双端阻断过氧化物酶模拟 DNA 酶对新生儿感染铜绿假单胞菌进行简单灵敏的检测。
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-05-20 DOI: 10.1080/07366205.2024.2348295
Dongyun Chen, Yicong Pan, Huan Yu, Xiaoxiang Chen
{"title":"Simple and sensitive detection of <i>Pseudomonas aeruginosa</i> in neonatal infection based on a both-end blocked peroxidase-mimicking DNAzyme.","authors":"Dongyun Chen, Yicong Pan, Huan Yu, Xiaoxiang Chen","doi":"10.1080/07366205.2024.2348295","DOIUrl":"https://doi.org/10.1080/07366205.2024.2348295","url":null,"abstract":"<p><p>Developing a simple and highly sensitive approach for <i>Pseudomonas aeruginosa</i> (<i>P. aeruginosa</i>) detection is crucial, as it is closely associated with various disorders, such as newborn infections. Nevertheless, few of techniques have the capability to accurately identify <i>P. aeruginosa</i> with a high level of sensitivity and significantly improved stability. The employment of the both-end blocked peroxidase-mimicking DNAzyme significantly diminished the interferences from background signals, so conferring the approach with a high degree of selectivity and reproducibility. The proposed method is demonstrated with exceptional discernment capacity in differentiating interfering microorganisms. The simplicity, elevated sensitivity and high discerning capability make the method a highly promising alternative instrument for pathogenic bacteria detection.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"76 7","pages":"323-332"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142054861","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
Cassava for the future: embryogenic liquid cultures suitable for new biotech techniques. 未来的木薯：适合新生物技术的胚胎液体培养物。
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-10-16 DOI: 10.1080/07366205.2024.2393546
Beata Dedičová, Luis Augusto Becerra Lopez-Lavalle
{"title":"Cassava for the future: embryogenic liquid cultures suitable for new biotech techniques.","authors":"Beata Dedičová, Luis Augusto Becerra Lopez-Lavalle","doi":"10.1080/07366205.2024.2393546","DOIUrl":"10.1080/07366205.2024.2393546","url":null,"abstract":"<p><p>Cassava, a crop of importance for subsistence farming in Africa, Asia, and Latin America, has the potential to benefit from global economic integration as a versatile industrial resource. Enhancing cassava productivity is not just a matter of agricultural competitiveness but a crucial step toward ensuring many communities' food security and livelihoods. Given its high performance in marginal environments, where climate change poses threats, ensuring food security and livelihoods relies on rapidly adapting cassava. This study aimed to develop a protocol that swiftly transitions cassava embryogenic short-period liquid suspension cultures, facilitating the regeneration of genetically stable <i>in vitro</i> plants. The resulting protocol, with its potential to be a foundational component in future technologies employing various genome editing or genetic modification techniques, holds promise for the advancement of cassava biotechnology.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"453-461"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142457179","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 standardized protocol for sample preparation for scanning electron microscopy to visualize extrachromosomal DNA. 用于扫描电子显微镜观察染色体外 DNA 的样本制备标准化方案。
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-06-19 DOI: 10.1080/07366205.2024.2346042
Jillann A Madren, Jingting Chen, William Dennis, Christina Ford, Kristen K White, Elizabeth Brunk
{"title":"A standardized protocol for sample preparation for scanning electron microscopy to visualize extrachromosomal DNA.","authors":"Jillann A Madren, Jingting Chen, William Dennis, Christina Ford, Kristen K White, Elizabeth Brunk","doi":"10.1080/07366205.2024.2346042","DOIUrl":"https://doi.org/10.1080/07366205.2024.2346042","url":null,"abstract":"<p><p>Extrachromosomal DNA (ecDNA) are circular DNA structures associated with cancer and drug resistance. One specific type, double minute (DM) chromosomes, has been studied since the 1960s using imaging techniques like cytogenetics and fluorescence microscopy. Specialized techniques such as atomic force microscopy (AFM) and scanning electron microscopy (SEM) offer micro to nano-scale visualization, but current sample preparation methods may not optimally preserve ecDNA structure. Our study introduces a systematic protocol using SEM for high-resolution ecDNA visualization. We have optimized the end-to-end procedure, providing a standardized approach to explore the circular architecture of ecDNA and address the urgent need for better understanding in cancer research.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"76 7","pages":"311-321"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142054859","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
Integrity of RNA in long-term-stored cervical liquid-based cytology samples: implications for biomarker research. 长期储存的宫颈液基细胞学样本中 RNA 的完整性：对生物标记研究的影响。
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-05-01 DOI: 10.2144/btn-2023-0112
Madeleine P J White, Andrew Stevenson, Hana Elasifer, Chris Davies, Kyriaki Nomikou, Kate Cuschieri, Sheila V Graham
{"title":"Integrity of RNA in long-term-stored cervical liquid-based cytology samples: implications for biomarker research.","authors":"Madeleine P J White, Andrew Stevenson, Hana Elasifer, Chris Davies, Kyriaki Nomikou, Kate Cuschieri, Sheila V Graham","doi":"10.2144/btn-2023-0112","DOIUrl":"10.2144/btn-2023-0112","url":null,"abstract":"<p><p>Biobanks of cervical screening (LBC) samples annotated with disease status are an invaluable resource to support the development of tools for the risk stratification of disease. Although there is growing interest in the assessment of RNA-based biomarkers, little is known on the suitability and durability of stored clinical samples (commonly used in cervical screening) to support RNA-based research. RNA was extracted from 260 stored LBC samples. Storage at -80°C or -25°C allowed isolation of sufficient RNA for further analysis. RNA was found to be substantially degraded according to Agilent Bioanalyser data. Despite this, RT-qPCR was successful in 95% samples tested. These data suggest that biobanked LBC samples are suitable for RNA-based assessment even if stored for up to 14 years.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"245-253"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140852647","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
Novel buffer for long-term preservation of DNA in biological material at room temperature. 用于在室温下长期保存生物材料中 DNA 的新型缓冲液。
IF 2.2 4区 工程技术
BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-07-01 DOI: 10.1080/07366205.2024.2360813
Mohaimin Kasu, Peter G Ristow, Adria Michelle Burrows, Zafrir Kuplik, Mark J Gibbons, Maria E D'Amato
{"title":"Novel buffer for long-term preservation of DNA in biological material at room temperature.","authors":"Mohaimin Kasu, Peter G Ristow, Adria Michelle Burrows, Zafrir Kuplik, Mark J Gibbons, Maria E D'Amato","doi":"10.1080/07366205.2024.2360813","DOIUrl":"10.1080/07366205.2024.2360813","url":null,"abstract":"<p><p>The collection and preservation of biological material before DNA analysis is critical for inter alia biomedical research, medical diagnostics, forensics and biodiversity conservation. In this study, we evaluate an in-house formulated buffer called the Forensic DNA Laboratory-buffer (FDL-buffer) for preservation of biological material for long term at room temperature. Human saliva stored in the buffer for 8 years, human blood stored for 3 years and delicate animal tissues from the jellyfish <i>Pelagia noctiluca</i> comb jelly <i>Beroe</i> sp., stored for 4 and 6 years respectively consistently produced high-quality DNA. FDL-buffer exhibited compatibility with standard organic, salting out and spin-column extraction methods, making it versatile and applicable to a wide range of applications, including automation.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"357-370"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141466005","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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