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Recording the brain in vivo: emerging technologies for the exploration of mental health conditions. Biotechniques (IF 2.7) Pub Date : 2024-03-14 Tristan Free
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 in vivo. [Formula: see text].
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Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d. Biotechniques (IF 2.7) Pub Date : 2024-02-29 Joaquin Abugattas-Nuñez Del Prado, Yi Ding, Jan de Sonneville, Kees-Jan van der Kolk, Miguel A Moreno-Mateos, Edward Málaga-Trillo, Herman P Spaink
In this study, the authors compared the efficiency of automated robotic and manual injection methods for the CRISPR-RfxCas13d (CasRx) system for mRNA knockdown and Cas9-mediated DNA targeting in zebrafish embryos. They targeted the no tail (TBXTA) gene as a proof-of-principle, evaluating the induced embryonic phenotypes. Both Cas9 and CasRx systems caused loss of function phenotypes for TBXTA. Cas9
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A free and user-friendly software protocol for the quantification of microfauna swimming behavior. Biotechniques (IF 2.7) Pub Date : 2024-02-29 Colleen M McMaken, Kristin E Gribble
Characterizing swimming behavior can provide a holistic assessment of the health, physiology and ecology of microfaunal species when done in conjunction with measuring other biological parameters. However, tracking and quantifying microfauna swimming behavior using existing automated tools is often difficult due to the animals' small size or transparency, or because of the high cost, expertise, or
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Multiparameter screen optimizes immunoprecipitation. Biotechniques (IF 2.7) Pub Date : 2024-02-29 Shaoshuai Xie, Leila Saba, Hua Jiang, Omar R Bringas, Mehrnoosh Oghbaie, Luciano Di Stefano, Vadim Sherman, John LaCava
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
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Considerations for developing mitochondrial transplantation techniques for individualized medicine. Biotechniques (IF 2.7) Pub Date : 2024-02-29 Kit Neikirk, Dominique C Stephens, Heather K Beasley, Andrea G Marshall, Jennifer A Gaddy, Steven M Damo, Antentor O Hinton
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.
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Age is just a number: can proteomic analysis provide insight into the aging body? Biotechniques (IF 2.7) Pub Date : 2024-02-22 Jenny Straiton
Recent proteomic studies have increased our understanding of the molecular process of aging, potentially enabling age-related diseases to be treated before the appearance of symptoms. [Formula: see text].
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An industry perspective approach and control strategy for implementation of ready-to-use cells in bioassays: survey outcome and recommendations. Biotechniques (IF 2.7) Pub Date : 2024-02-09 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
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
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Extracting high-quality RNA from formaldehyde-fixed naturally aged neuromusculoskeletal tissues. Biotechniques (IF 2.7) Pub Date : 2024-02-09 Seth D Thompson, Rajeswari Pichika, Richard L Lieber, Mitra Lavasani
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
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A novel method for sampling subgingival microbiome: a comparative metatranscriptomic study. Biotechniques (IF 2.7) Pub Date : 2024-02-06 Diana Demusaj, Ryan Toma, Tanveer Khan, Lan Hu, Guruduth Banavar, Momchilo Vuyisich
The subgingival microbiome has been implicated in oral and systemic diseases such as periodontitis and Alzheimer's disease. However, subgingival sampling is challenging. We developed a novel method of sampling the subgingival microbiome by rotationally swabbing the supragingival area, named subgingival-P (for proxy) samples. We sampled and metatranscriptomically analyzed subgingival and subgingival-P
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High-throughput screening for a SARS-CoV-2 frameshifting inhibitor using a cell-free protein synthesis system. Biotechniques (IF 2.7) Pub Date : 2024-01-31 Kodai Machida, Rin Tanaka, Seraya Miki, Shotaro Noseda, Mayumi Yuasa-Sunagawa, Hiroaki Imataka
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
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Assessment of measurement accuracy of amplified DNA using a colorimetric loop-mediated isothermal amplification assay. Biotechniques (IF 2.7) Pub Date : 2023-12-22 Seongsoo Hong, Jeongho Park, Jaekyung Bok, Euna Cho, Joowon Rhee
A colorimetric loop-mediated isothermal amplification assay detects changes in pH during amplification based on color changes at a constant temperature. Currently, various studies have focused on developing and assessing molecular point-of-care testing instruments. In this study, we evaluated amplified DNA concentrations measured using the colorimetric LAMP assay of the 1POT™ Professional device (1drop
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An optimal genomic DNA extraction method for shoots of four Dendrocalamus species based on membership function analysis. Biotechniques (IF 2.7) Pub Date : 2023-12-22 Yingchun Ma, Bowen Li, Dejia Yang, Shuguang Wang, Lixia Yu, Hui Zhan, Juan Li
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 Dendrocalamus 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
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Reference genes for gene expression profiling in mouse models of Listeria monocytogenes infection. Biotechniques (IF 2.7) Pub Date : 2023-12-19 Lethicia Souza Tavares, Roberta Lane Oliveira-Silva, Marcelo Tigre Moura, Jéssica Barboza da Silva, Ana Maria Benko-Iseppon, José Vitor Lima-Filho
RT-qPCR dissects transcription-based processes but requires reference genes (RGs) for data normalization. This study prospected RGs for mouse macrophages (pMØ) and spleen infected with Listeria monocytogenes. The pMØ were infected in vitro with L. monocytogenes or vehicle for 4 h. Mice were injected with L. monocytogenes (or vehicle) and euthanized 24 h post-injection. The RGs came from a multispecies
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Comparison of colorimetric, spectroscopic and electrochemical techniques for quantification of hydrogen sulfide. Biotechniques (IF 2.7) Pub Date : 2023-12-07 Susmit Mhatre, Anjali Rai, Hatim Ali, Akash Patil, Neetu Singh, Richa Verma, John Auden, Cole Chandler, Alekha Dash, Catherine Opere, Somnath Singh
Background: Hydrogen sulfide (H2S), an endogenous gasotransmitter, has potential applications in several conditions. However, its quantification in simulated physiological solutions is a major challenge due to its gaseous nature and other physicochemical properties. Aim: This study was designed to compare four commonly used H2S detection and quantification methods in aqueous solutions. Methods: The
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Cancer's sweet spot: techniques to harness saccharides in tumor biology. Biotechniques (IF 2.7) Pub Date : 2023-12-04 Jade Parker
All of the cells in our bodies are enveloped in sugar, this sweet coating plays a particularly interesting and crucial role in tumor biology. Here, we review the techniques being used to detect and exploit cancer's sweet spot. including click chemistry, glycoproteomic profiling and bioorthogonal chemistry.
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Automated method for quantification of 20 amino acids in cell culture media during biopharmaceutical development. Biotechniques (IF 2.7) Pub Date : 2023-11-24 Ankita Dubey, Srishti Joshi, Kratika Upadhyay, Ansuman Mahato, Anurag S Rathore
Herein, a step-by-step protocol for simultaneous detection of 20 amino acids commonly present in cell culture media is described. The protocol facilitates detection of both primary and secondary amino acids through a two-step precolumn derivatization strategy using ortho-phthalaldehyde and 9-fluorenylmethyl chloroformate as derivatizing agents. The separation of derivatized amino acids with varying
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PS4: a next-generation dataset for protein single-sequence secondary structure prediction. Biotechniques (IF 2.7) Pub Date : 2023-11-24 Omar Peracha
Protein secondary structure prediction is a subproblem of protein folding. A light-weight algorithm capable of accurately predicting secondary structure from only the protein residue sequence could provide useful input for tertiary structure prediction, alleviating the reliance on multiple sequence alignments typically seen in today's best-performing models. Unfortunately, existing datasets for secondary
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Three-dimensional visualization and analysis of dendritic spines in human brain tissue. Biotechniques (IF 2.7) Pub Date : 2023-11-23 Haitao Sun, Hei Ming Lai, Wutian Wu
We developed a simple yet powerful technique to visualize neuronal morphology in human brain tissues. By ballistically shooting DiI (1,1'-dioctadecyl-3,3,3'3'-tetramethylindocarbocyanine perchlorate)-coated tungsten particles to randomly label neurons, then clearing tissues with OPTIClear, we demonstrated the tracing of branched dendritic trees and spines in three dimensions. High-resolution imaging
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Decoding disease: Scott Patterson's perspectives on the power of biomarkers in drug development. Biotechniques (IF 2.7) Pub Date : 2023-11-23 Scott D Patterson
Scott Patterson (Gilead Sciences Inc., CA, USA) speaks to Ashling Cannon, Journal Development Editor at BioTechniques, about his career. Patterson is a biochemist and proteomics and biomarker/translational expert with over 30 years of industry experience following 13 years in an academic setting. Patterson earned his BSc and PhD in Physiology and Pharmacology from the University of Queensland (Australia)
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Cradle cultures: growing stem cell-derived developmental cell models in vitro. Biotechniques (IF 2.7) Pub Date : 2023-11-16 Beatrice Bowlby
How are three stem cell-derived developmental cell models furthering our understanding of post-implantation human embryo development? And why have recent advancements in these human embryo-like models spurred ethical discussion and the need to refine our definition of 'embryo'? [Formula: see text].
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Predicting molecular docking of per- and polyfluoroalkyl substances to blood protein using generative artificial intelligence algorithm DiffDock. Biotechniques (IF 2.7) Pub Date : 2023-11-10 Dhan Lord B Fortela, Ashley P Mikolajczyk, Miranda R Carnes, Wayne Sharp, Emmanuel Revellame, Rafael Hernandez, William E Holmes, Mark E Zappi
This study computationally evaluates the molecular docking affinity of various perfluoroalkyl and polyfluoroalkyl substances (PFAs) towards blood proteins using a generative machine-learning algorithm, DiffDock, specialized in protein-ligand blind-docking learning and prediction. Concerns about the chemical pathways and accumulation of PFAs in the environment and eventually in the human body has been
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Considerations for colorblind individuals on selecting colorimetric or fluorescent dye assay outcomes. Biotechniques (IF 2.7) Pub Date : 2023-11-09 Kirstyn Loyva, Erik Hofmeister, Fiona Georgousi, Constance Roderick, Rebecca Cole
A disadvantage of colorimetric detection in nucleic acid amplification assays is the possibility that a colorblind individual may interpret colors differently than observers with full-color vision. Using an isothermal amplification assay, the ability of colorblind individuals to distinguish between positive and negative results for four dyes was tested. Five individuals with self-reported colorblindness
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MSCProfiler: a single cell image processing workflow to investigate mesenchymal stem cell heterogeneity. Biotechniques (IF 2.7) Pub Date : 2023-11-02 Ayona Gupta,Safia Kousar Shaik,Lakshmi Balasubramanian,Uttara Chakraborty
Single cell cytometry has demonstrated plausible immuno-heterogeneity of mesenchymal stem cells (MSCs) owing to their multivariate stromal origin. To contribute successfully to next-generation stem cell therapeutics, a deeper understanding of their cellular morphology and immunophenotype is important. In this study, the authors describe MSCProfiler, an image analysis pipeline developed using CellProfiler
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A simple, sensitive and colorimetric assay for Pseudomonas aeruginosa infection analysis. Biotechniques (IF 2.7) Pub Date : 2023-10-26 Jie Liu,Dan Lu,Junyuan Wang
Skin and soft tissue infections caused by Pseudomonas aeruginosa are common acquired diseases in postpartum care. Many methods have been developed in recent years for detecting P. aeruginosa, but they are criticized for the drawbacks of labor-intensiveness, complicated operation and high cost. Here, a simple, sensitive and colorimetric assay for P. aeruginosa detection is described. The approach displays
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An easy method for quantification of anaerobic and microaerobic gene expression with fluorescent reporter proteins. Biotechniques (IF 2.7) Pub Date : 2023-10-26 Lucas Pedraz,Eduard Torrents
Fluorescent proteins, such as green fluorescent proteins, are invaluable tools for detecting and quantifying gene expression in high-throughput reporter gene assays. However, they introduce significant inaccuracies in studies involving microaerobiosis or anaerobiosis, as oxygen is required for the maturation of these proteins' chromophores. In this study, the authors highlight the errors incurred by
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A simple and rapid method for isolating high-quality RNA from kenaf with high polysaccharide and polyphenol contents. Biotechniques (IF 2.7) Pub Date : 2023-10-26 Xiaofang Liao,Yanhong Zhao,Hongwei Li,Wenhuan Hou,Xingfu Tang,Ruiyang Zhou
The extraction of high-quality RNA from kenaf is essential for genetic and molecular biology research. However, the presence of high levels of polysaccharide and polyphenol compounds in kenaf poses challenges for RNA isolation. We proposed a simplified, time-saving and cost-effective method for isolating high quantities of RNA from various kenaf tissues. This method exhibited superior efficiency in
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Decoding the mechanisms of embryo development with spatial biology. Biotechniques (IF 2.7) Pub Date : 2023-10-19 Asisha Al-Janabi
[Formula: see text] Spatial genomics first emerged in the 1990s to map biological function data onto multidimensional images of embryo specimens and has continued to be instrumental in studying embryogenesis and understanding early human development in the decades since.
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A simplified and rapid in situ hybridization protocol for planarians. Biotechniques (IF 2.7) Pub Date : 2023-10-18 Andrew J Gaetano, Ryan S King
Whole-mount in situ hybridization is a critical technique for analyzing gene expression in planarians. While robust in situ protocols have been developed, these protocols are laborious, making them challenging to incorporate in an academic setting, reducing throughput and increasing time to results. Here, the authors systematically tested modifications to all phases of the protocol with the goal of
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Generation of UL128-shRNA transduced fibroblasts for the release of cell-free virus from clinical human cytomegalovirus isolates. Biotechniques (IF 2.7) Pub Date : 2023-10-17 Nina Weiler, Kerstin Laib Sampaio, Myriam Scherer, Christian Sinzger
Working with recent isolates of human cytomegalovirus (HCMV) is complicated by their strictly cell-associated growth with lack of infectivity in the supernatant. Adaptation to cell-free growth is associated with disruption of the viral UL128 gene locus. The authors transduced fibroblasts with a lentiviral vector encoding UL128-specific-shRNA to allow the release of cell-free infectivity without genetic
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PlayBack cloning: simple, reversible, cost-effective cloning for the combinatorial assembly of complex expression constructs. Biotechniques (IF 2.7) Pub Date : 2023-10-10 Gregory Foran, Ryan D Hallam, Marvel Megaly, Anel Turgambayeva, Aleksandar Necakov
With advancements in multicomponent molecular biological tools, the need for versatile, rapid and cost-effective cloning that enables successful combinatorial assembly of DNA plasmids of interest is becoming increasingly important. Unfortunately, current cloning platforms fall short regarding affordability, ease of combinatorial assembly and, above all, the ability to iteratively remove individual
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Single-cell RNA sequencing of bronchoscopy specimens: development of a rapid minimal handling protocol. Biotechniques (IF 2.7) Pub Date : 2023-10-10 Firoozeh V Gerayeli, Stephen Milne, Chen Xi Yang, Xuan Li, Elizabeth Guinto, Julia Shun-Wei Yang, Chung Yan Cheung, Tara R Stach, Tawimas Shaipanich, Janice M Leung, Don D Sin
Single-cell RNA sequencing (scRNA-seq) is an important tool for understanding disease pathophysiology, including airway diseases. Currently, the majority of scRNA-seq studies in airway diseases have used invasive methods (airway biopsy, surgical resection), which carry inherent risks and thus present a major limitation to scRNA-seq investigation of airway pathobiology. Bronchial brushing, where the
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Microbial forensics: what we've learned from Amerithrax and beyond. Biotechniques (IF 2.7) Pub Date : 2023-10-06 Louis Gautier
The Amerithrax investigation into anthrax letter attacks in the USA forever changed the game in microbial forensics. Here we review the techniques used, then and now, to neutralize bioterrorism threats. [Formula: see text].
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A practical approach to genome assembly and annotation of Basidiomycota using the example of Armillaria. Biotechniques (IF 2.7) Pub Date : 2023-09-08 Deborah L Narh Mensah, Brenda D Wingfield, Martin Pa Coetzee
Technological advancements in genome sequencing, assembly and annotation platforms and algorithms that resulted in several genomic studies have created an opportunity to further our understanding of the biology of phytopathogens, including Armillaria species. Most Armillaria species are facultative necrotrophs that cause root- and stem-rot, usually on woody plants, significantly impacting agriculture
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A cell hybridization-based method of generating recombinant rabbit monoclonal antibodies for detecting cytokines. Biotechniques (IF 2.7) Pub Date : 2023-09-06 Yu Peng, Chun'e Zhang, Minyan Deng, Haijuan Jiang, Huishu Huang, Yue Li, Weiping Lai, Yu-Pin Lin, Jun Yu
Recombinant rabbit monoclonal antibodies (rabbit rAbs) have shown promise in various biomedical fields. However, it is challenging and costly to generate rabbit rAbs using traditional techniques. Here we describe a convenient and cost-effective method. Using this method, we generated rabbit rAbs against mouse soluble IL-6 receptor α with affinities in the range of 10-9 to 10-12 M. The presented method
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Special Focus Issue - Celebrating 40 years of publishing lab techniques: looking back to look foreword. Biotechniques (IF 2.7) Pub Date : 2023-09-01 Beatrice Bowlby,Tristan Free,Annie Coulson,Aisha Al-Janabi,Ashling Cannon,Ebony Torrington
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Education in microscopy: taking a closer look with Jennifer Waters. Biotechniques (IF 2.7) Pub Date : 2023-08-29 Jennifer Waters
Jennifer Waters (Nikon Imaging Center, Harvard Medical School, MA, USA) spoke to Ebony Torrington (Future Science Group, London, UK) about her career in microscopy and the importance of education. Jennifer is the Director of the Nikon Imaging Center (NIC) at Harvard Medical School and a Chan Zuckerberg Initiative (CZI) Imaging Scientist. She was trained in cell biology and microscopy by Ted Salmon
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Unraveling crime scenes strand by strand: the forensic odyssey of Bruce Budowle. Biotechniques (IF 2.7) Pub Date : 2023-08-25 Bruce Budowle
Bruce Budowle speaks to Ashling Cannon, Journal Development Editor for BioTechniques, about advancements & challenges in forensic science. Budowle completed his doctorate in genetics at Virginia Tech (VA, USA) formally known as Virginia Polytechnic Institute and State University. He then went on to complete a postdoctoral fellowship at the University of Alabama at Birmingham (AL, USA) to study genetic
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I know what you're thinking; can neuroimaging truly reveal our innermost thoughts? Biotechniques (IF 2.7) Pub Date : 2023-08-25 Jenny Straiton
[Formula: see text] Advances in neuroimaging, combined with developments in artificial intelligence software, have allowed researchers to non-invasively decode the brain and 'read the mind'.
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Colorimetric analysis of C-reactive protein via 'jellyfish' probe-based exonuclease III-assisted multiple-signal recycles. Biotechniques (IF 2.7) Pub Date : 2023-08-24 Jinghui An, Huajun Wang, Tong Su, Fengwu Shi, Su Liu
C-reactive protein (CRP) is a potential biomarker for evaluating inflammatory responses in patients receiving coronary artery bypass graft surgery. Here, the authors depict a sensitive and reliable colorimetric approach for CRP analysis. In this method, an aptamer specifically binds with CRP and an initiator sequence is released from an arch probe to activate signal amplification. The released initiator
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Quantitative analysis of lncRNA in formalin-fixed paraffin-embedded tissues of oral squamous cell carcinoma. Biotechniques (IF 2.7) Pub Date : 2023-08-17 Kiran Kumar,Kaveri Hallikeri,Ajaykumar Oli,Mallikarjun Goni,Apoorva Jain,Jagadeesha Poyya,Alagilavada S Shilpasree,Palaksha Kanive Javaregowda
The study evaluated expression profiles of few regulatory lncRNAs in oral squamous cell carcinoma and normal mucosa adjacent to oral cancer using paired fresh frozen and formalin-fixed paraffin-embedded (FFPE) tissues stored at a different duration of time (1-5 years) using real-time quantitative PCR. The quantity and quality of total RNA isolated from FFPE tissues was less compared with that of fresh
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Peter and the Beanstalk: tackling the giant questions of soybean nodulation. Biotechniques (IF 2.7) Pub Date : 2023-08-17 Peter Gresshoff
The following is an edited interview, carried out by the Journal Development Editor of BioTechniques, Ashling Cannon, with Peter Gresshoff (University of Queensland, UQ; Brisbane, Australia). Peter is a plant developmental geneticist, using molecular and genetic tools to understand the complexities of gene networks during the control of nodule formation in legumes. He was the Director of the Center
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Releasing secrets bound to ancient remains with modern DNA extraction techniques: an interview with Elena Essel. Biotechniques (IF 2.7) Pub Date : 2023-08-17 Elena Essel
Elena Essel (Msc) spoke to Ebony Torrington, Managing Editor of BioTechniques. 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
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A high-throughput pipeline for DNA/RNA/small RNA purification from tissue samples for sequencing. Biotechniques (IF 2.7) Pub Date : 2023-08-08 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
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.
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Selective expansion of target cells using the Enrich TROVO platform. Biotechniques (IF 2.7) Pub Date : 2023-08-08 Prem Shrestha, Stephen Rotarori, Kirby Madden-Hennessey, Christina Mohammed, Chi-Han Yang, Jordan Urbani, Joseph Pettinelli, Xueqi Liu, Qi Zhao
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
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HIV: the pursuit of an elusive vaccine. Biotechniques (IF 2.7) Pub Date : 2023-08-04 Annie Coulson
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].
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A novel use of a needle-free injection system for improved nucleic acid delivery and expression in vivo. Biotechniques (IF 2.7) Pub Date : 2023-07-27 Nathan Liu,Hani Abd-Ul-Salam,Noémie Joannette-Lafrance,Jingjing Li,Karim Menassa,Monzur Murshed
Transfection, a nonviral method of nucleic acid delivery, often exhibits poor efficiency in vivo. The needle-based in vivo delivery of transfection reagents can be invasive. Here, we report a noninvasive protocol for in vivo 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
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The root microbiome: techniques for exploration and agricultural applications. Biotechniques (IF 2.7) Pub Date : 2023-07-26 Ashling Cannon
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].
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Overlap extension cloning of PCR products into a Gateway-compatible plasmid vector. Biotechniques (IF 2.7) Pub Date : 2023-07-09 Omid Zare-Mehrjerdi,Gracie Trader,Viktor Kirik
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
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Chromatin immunoprecipitation and library preparation: a powerful tool to unravel the epigenome. Biotechniques (IF 2.7) Pub Date : 2023-06-29 Mukesh Pratap Yadav,Vikas Kundra
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Microbial marvels: could microbes resolve climate change? Biotechniques (IF 2.7) Pub Date : 2023-06-24 Jade Parker
Our planet faces many challenges. Namely, the damage caused by human disruption of natural levels of carbon and nitrogen as a result of the use of fossil fuels. Microbes play a crucial role in the biogeochemical processing of these elements, in turn placing them at the heart of one of the most testing issues of all time - climate change. [Formula: see text].
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PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNA. Biotechniques (IF 2.7) Pub Date : 2023-06-21 Yasunori Nishida,Kotetsu Kayama,Taichi Endoh,Kiwamu Hanazono,Gerry Amor Camer,Daiji Endoh
We formulated a method to synthesize 1 kbp DNA fragments using 'oligomer unidirectional joining method' via asymmetric extension supported by a simulator for oligonucleotide extension (AESOE). In this study, trials were conducted on 41 sets of different genomic pieces of ten flaviviral genomes, and 31 bacterial 16s rRNA fragments with sizes ranging from 500 bases to 1.0 kbp. Synthetic gene production
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Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technology. Biotechniques (IF 2.7) Pub Date : 2023-06-21 Masako Osawa,Takurou Akiya,Funa Ogawa,Takao Suzuki,Masaki Yamagami,Tadashi Umemoto,Akira Ideno
Quantitative bioanalysis is essential when establishing pharmacokinetic properties during the drug development process. To overcome challenges of sensitivity, specificity and process complexity associated with the conventional analysis of antisense oligonucleotides (ASOs), a new approach to nonenzymatic hybridization assays using probe alteration-linked self-assembly reaction (PALSAR) technology as
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Multiscale computational modeling offers key to understanding molecular logic underpinning development and disease. Biotechniques (IF 2.7) Pub Date : 2023-06-16 Himanshu Kaul
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The path to solving the protein folding problem. Biotechniques (IF 2.7) Pub Date : 2023-6-12 Jenny Straiton
[Formula: see text] With advances in imaging technologies and the development of artificial intelligence-based predictive software, has the protein folding problem finally been solved?
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Developing virtual and augmented reality applications for science, technology, engineering and math education. Biotechniques (IF 2.7) Pub Date : 2023-06-09 Christopher L Hemme,Rachel Carley,Arielle Norton,Moez Ghumman,Hannah Nguyen,Ryan Ivone,Jyothi U Menon,Jie Shen,Matthew Bertin,Roberta King,Elizabeth Leibovitz,Roy Bergstrom,Bongsup Cho
The Rhode Island IDeA Network of Biomedical Research Excellence Molecular Informatics Core at the University of Rhode Island Information Technology Services Innovative Learning Technologies developed virtual and augmented reality applications to teach concepts in biomedical science, including pharmacology, medicinal chemistry, cell culture and nanotechnology. The apps were developed as full virtual
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A modified method for efficient RNA isolation from mangrove root tissues rich in secondary metabolites. Biotechniques (IF 2.7) Pub Date : 2023-06-08 Ashifa Nizam,Haritha Kalath,Ajay Kumar
Secondary metabolites in mangroves often interfere with RNA extraction yielding poor concentration and quality, which is unsuitable for downstream applications. As existing protocols yielded low-quality RNA from root tissues of Kandelia candel (L.) Druce and Rhizophora mucronata Lam., an optimized method was developed for improving the quality and yield of RNA. Compared with three other methods, this
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Site-specific analysis of ribosomal 2'O-methylation by quantitative reverse transcription PCR under low deoxynucleotide triphosphate concentrations. Biotechniques (IF 2.7) Pub Date : 2023-06-05 Daniela Barros-Silva,Johan Tsui,Carmen Jerónimo,Guido Jenster,Elena S Martens-Uzunova
Ribose 2'O-methylation (Nm, ribomethylation) is the most abundant RNA modification present in rRNA. It has been shown that alterations in ribosomal 2'O-methylation at individual Nm sites likely reflect regulated cellular processes. Although several analytical approaches for Nm detection and profiling have been developed, a simple and affordable method for the screening and measurement of individual
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Direct-current electric field stimulation promotes proliferation and maintains stemness of mesenchymal stem cells. Biotechniques (IF 2.7) Pub Date : 2023-06-01 Mengchang Liu,Defu Xie,Huizhen Zeng,Ning Zhai,Lan Liu,Hong Yan
Mesenchymal stem cells are frequently utilized in the study of regenerative medicine. Electric fields (EFs) influence many biological processes, such as cell proliferation, migration and differentiation. In the present study, a novel device capable of delivering a direct current of EF stimulation to cells cultured in vitro is described. This bioreactor was customized to simultaneously apply a direct-current
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Gibson assembly interposition improves amplification efficiency of long DNA and multifragment overlap extension PCR. Biotechniques (IF 2.7) Pub Date : 2023-05-31 Junyi Liu,Fangyin Liu,Xueer Luo,Ming Chen,Chengjun Wang,Liuyue Wang,Huabo Chen
For difficult overlap extension PCR, a Gibson assembly process was inserted between the two PCR rounds to facilitate the formation of complete gene templates at a moderate temperature. That is, after amplifying each DNA fragment, they were preluded by a Gibson assembly process in equal proportion. Then, the assembled mixture was used as a template for the second PCR round. This idea was tested and
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An optimized TRIzol-based method for isolating RNA from adipose tissue. Biotechniques (IF 2.7) Pub Date : 2023-05-26 Hongwei Zhang,Yaoming Liu,Bingcheng Yu,Rong Lu
High-quality RNA isolation from recalcitrant adipose tissue with high lipid content and low cell numbers is difficult. Many studies have made efforts to optimize methods for isolating RNA from adipose tissue through combinations of column-based kits and phenol-chloroform methods, or through in-house protocols. However, the considerable complexity of these protocols and the various kits/materials required