... standard EGFP plasmid that used to be sold by Clontech and is in a freezer box in just about every lab...environmental changes that the biosensor is designed to detect resulting in false positives [2, 4]. Solutions ...molecular sizing columns and native gels fail to detect EGFP dimerization. Importantly, all of these assays...assays are performed in vitro. Assays to detect FP dimerization in in vivo cellular contexts, such as ... from nonbioluminescent Anthozoa species. Nat Biotechnol, 1999. 17(10): p. 969-73. PubMed PMID: 10504696...

...To achieve this, scientists use a multitude of techniques including co-transfection of two or more plasmids...started with them. Why Use multicistronic vectors? Detecting cells that are expressing your gene, especially...straightforward process. Rather than try to directly detect your gene of interest, scientists have instead ...not have to exclusively be used as a means of detection; they are useful almost anytime you want to express...upstream of an IRES fluorescent reporter for easy detection.5  Just released: Addgene's Plasmids 101 eBook...

... last 5 years to incorporate more science and technology into the school curriculum. Our Parent Teacher...parents who can answer some of the surprisingly technical questions elementary students can ask. It doesn...and the internet before it, genome engineering technology is moving along much faster than it can be officially...sugarcoat the potential dangers of people using the technology in an unethical manner, but I also tried not ... timely.” Genome engineering, and many other technologies and fields in biology, are moving so quickly...

...between neighboring fluorophores, FRET can be used to detect the inter- and intramolecular interactions that... fluorescence complementation is a method for detecting protein-protein interactions that uses split fluorescent...FP results in a fluorescent signal that can be detected by fluorescence microscopy. Figure 1: How...Medical Center, where he used molecular biology techniques to study inflammation and neurodegeneration. ...BiFC): A 5-year update and future perspectives. Biotech 53. https://doi.org/10.2144/000113943 Additional...

...ELISAs. Immunofluorescence. What do all of these techniques have in common? They all typically require secondary.... Could there be an alternative method for immunodetection? Enter the Görlich lab and their anti-mouse...antibodies for all of the above listed molecular techniques, but they can also be expressed in bacteria and...main reasons antibodies are great indirect immunodetection tools is their strong signal strength. This...Although antibodies have been the titans of immunodetection, nanobodies may be better suited for the job...

...recombinant Erythropoietin. J Biotechnol. 2020 Feb 10;309:44-52. doi: 10.1016/j.jbiotec.2019.12.017. Epub 2019...conformational changes in the antigen or lower detection sensitivity of the assay. Where possible, it can...beads from very high cell density suspension. Biotechnol Bioeng. 2021 Sep;118(9):3499-3510. doi: 10.1002...Roberts CJ. Protein aggregation - Mechanisms, detection, and control. Int J Pharm. 2018 Oct 25;550(1-2...

...Healthcare consultants advise pharmaceutical companies, biotech start-ups, health insurance companies, and governments...strategy tend to be mostly engaged by pharma and biotech clients. In this realm, there are a few different...organizations, investment and equity research firms, biotech start-ups, venture capital firms, and more. Essentially...science, with a newfound appreciation for how technologies get developed and commercialized after they ...healthcare industry publications like FiercePharma, FierceBiotech, and BioCentury can be an engaging way to learn...

...specific protein out of a complex mix. Using this technique, users can look for the presence or absence of...acids. Read on to learn more about this versatile technique.  Immunoprecipitation Overview An immunoprecipitation...species as the antibody that will be used for detection in a subsequent Western blot. When choosing beads...application. Typically, if the target protein will be detected in a Western blot then it can be eluted by boiling...epigenetics routinely use the chromatin IP (ChIP) technique to identify DNA binding proteins involved in histone...

...Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the absence of Cre. ... are determined using a chromogenic endotoxin detection assay based on the amebocyte lysate method. Purity.... mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from Karl Deisseroth (Addgene...for mycoplasma contamination using mycoplasma detection kits. Cell line is maintained for ~20 passages...

...Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306 Koornneef...protein-protein interactions in proximity labeling techniques such as BioID (Roux et al., 2012) and APEX (Rhee...tools, such as gene manipulation and mutagenesis techniques, are available to the scientific community.  ... as a Model Organism and Its Application in Biotechnology. In Escherichia coli - Recent Advances on Physiology...

...RNA production can be detected by using a MS2 coat protein that specifically detects a specific RNA stem...instance, pleckstrin homology domains can be used to detect specific phosphoinositides (Varnai and Balla, 2008...Cas9 is a protein, a fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP...fluorescent proteins (Chernov et al., 2017) may improve detection. Since longer wavelengths have better tissue penetration...

... (reviewed in Shcherbakova et al., 2018). For detection of protein-protein interactions, we recommend ...Multiplexing and Optogenetics across Scales. Trends Biotechnol 36, 1230-1243 (2018). Shcherbakova, D.M. et al...Sivanesan, D. & Michnick, S.W. An infrared reporter to detect spatiotemporal dynamics of protein-protein interactions...encoded calcium indicator for in vivo imaging. Nat Biotechnol 39, 368-377 (2021). Shcherbakova, D.M. Near-infrared... fluorescent protein for in vivo imaging. Nat Biotechnol 29, 757-761 (2011).   ...

...Site directed mutagenesis is a highly versatile technique that can be used to introduce specific nucleotide...naturally occuring SNPs in a plasmid context). The technique is also highly relevant in this age of CRISPR;...plasmid digestion (blue). Digestion with enzyme “B” detects primer multimerization (purple) which is evident...~3%). Note that primer duplication will evade detection in the initial RFLP analysis simply because the...on the Addgene Blog Brush up on Your Cloning Techniques Learn More about Plasmids in Our Plasmids 101...

...understanding the brain’s function is knowing its architecture, in particular the connections between different...nervous system With the advent of viral vector technology, new methods have been developed to study neuronal...consistent which may be caused by the diversity of techniques, species, and brain regions used. Evidence for... Gradinaru Lab at the California Institute of Technology generated novel AAVs (Chan et al., 2017) that...lower toxicity, higher specificity and reduced technical variability compared to chemical tracers. rAAV2...targets neonatal neurons and adult astrocytes. Nat Biotechnol 27, 59–65. PubMed PMID: 19098898. PubMed Central...

...authors use PhoCl in combination with this caging technique to beautifully demonstrate light-dependent Gal4... base editors The popular CRISPR base editing technique uses dCas9/Cas9 nickase fused to a cytidine deaminase... PubMed PMID: 28585549 Kim Y, et al. Nature Biotechnology. 2017. PubMed PMID: 28191901 Listen to Our ...a fully sequenced genome, many applications in biotech, and is considered the most studied Gram-positive...vivo. Utilizing CRISPR interference (CRISPRi) technology, the arrayed knockdown library targets 289 genes...In Narayan et al., they used this now classic technique to elucidate the mechanism by which SOX2 acts ...

...pathways. Trends in Biotechnology, 33(2), 92–100. https://doi.org/10.1016/j.tibtech.2014.11.007...activate the pathway than traditional pharmacology techniques allow. Multiple photoswitches can be used in ...reading Abreu, N., & Levitz, J. (2020). Optogenetic Techniques for Manipulating and Sensing G Protein-Coupled...References Abreu, N., & Levitz, J. (2020). Optogenetic Techniques for Manipulating and Sensing G Protein-Coupled...

...their hot-off-the-presses papers, the awesome new technology they’re utilizing, and the tricks they’ve used...plasmids in your own lab. Summaries of the latest technologies and breakthrough research. Insights about ..., post-doctoral fellow, lab manager, research technician, or Principal Investigator. How-to guides for...a special focus on cloning and plasmid-based technologies. Commentary about scientific sharing and the...

... Chapter 3 - Addgene as a leader in standardized tech transfer and the adoption of the UBMTA Chapter 4... and Diplomacy, with a focus on international technology management, from the Fletcher School at Tufts...investment banking analyst at Credit Suisse’s Technology Group in Palo Alto, CA.  Benjie Chen is a co-founder...and networks. Benjie was Addgene's Director of Technology for 7 years, and is now Chief Software Engineer...

...current forming membrane pores) transfection. These techniques are still used and efficiently transfer plasmid...not been tested for HIV.  If not using proper technique, there’s always a small chance that a researcher...transfection, CaCl2 transfection is a simple and cheap technique that should work well. Quickly after transfection...for virus titration: the p24 ELISA dosing method detects the expression of viral capsid antigen p24, the...Therapy, Dr. Ke Xu (Ed.), ISBN: 978-953-307-539-6, InTech. 4. Walter, W., and U. Stein. "Viral vectors for...

... itself was relatively easy. Figure 4: Detection of edited BRAF genomic loci in Cas9-expressing...dsDNA) at basepair mismatches. To use this tool to detect a BRAF gene edit, simply PCR-amplify the BRAF locus...be in the lab. Based on the power of available techniques and how quick and easy it is to share information...Sullender M, Ebert BL, Xavier RJ, Root DE. Nat Biotechnol. 2014 Sep 3. doi: 10.1038/nbt.3026. 10.1038/nbt...Orchard R, Virgin HW, Listgarten J, Root DE. Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. PubMed PMID...

...research.     Improved nucleic acid sequence detection Feng Zhang’s lab recently developed a platform...high-sensitivity enzymatic reporter unlocking) to detect single molecules of RNA or DNA. Now they have developed...developed this further to provide multiplexed detection, higher sensitivity, and more. Read the paper...and cleaves ssDNA which they have adapted for detecting SNPs. Learn more from this Addgene blog post...

... modular, standardized, disciplined, and open techniques. Chapter 4: Research projects underway in the...Synthetic Biology. Wide adoption of synthetic biology techniques in biology research as can already be seen with...University of Cambridge before spending time in biotech screening direct interactions between drugs and...investigates bacteria and extremophiles for biotechnology and has been directly involved in supervising...

...backgrounds! Show off your favorite reagent, scientific technique, or plasmid repository at your next meeting.   ...blog topic to learn more about FRET and other techniques using FPs! Otherwise, check out our plasmid and... browse plasmids by field, organism, research technique, and more. Download the green DNA background. ...those backgrounds representative of your favorite technique? Our lab-themed background is a great fit for ...

...depositors who quickly shared their reagents: detection methods, synthetic nanobodies, SARS-CoV-2 protein...reagent sharing even broader and began working with tech transfer offices to make reagents available to scientists...extracting RNA without a kit. 3. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas by Shravanti Suresh...University detailed five methods for using CRISPR to detect SARS-CoV-2, including many with plasmids deposited...

...classification system for the array of CRISPR technologies. Learning the general principles of this classification...systems. Type III systems are unique, as they can technically target both RNA and DNA, though it is largely...the first taught when introducing CRISPR as a technology. There are three subtypes, A–C, with the beloved...Y. (2018). Class 2 CRISPR/Cas: an expanding biotechnology toolbox for and beyond genome editing. Cell ...Yoshimi, K., & Mashimo, T. (2022b). Genome editing technology and applications with the type I CRISPR system...

...this post even more useful to your colleagues. Technique: Probe Phage Genomes for Host Binding Proteins...editor’s choice posts this month. The first, "Technique: Probe Phage Genomes for Host Binding Proteins...from the University of Alberta and describes a technique you can use to find phage proteins that bind to...2) The post describes an ingenious but simple technique to screen for proteins that could be useful for...

...are swarming to see what can be done with this technology to help patients. Dr. Stoffels and others in ...months compared to a year or greater with older technologies. Addgene is proud to be helping get CRISPR reagents..., the panelists agreed that, for example, the technology is already available today to cure sickle cell...really sophisticated”.     New to CRISPR/Cas9 Technology? Learn More: Addgene's CRISPR/Cas9 Guide History...

...It’s clear that CRISPR-Cas9 technology has really changed the game for anyone looking to quickly and ...question. Additionally, a concurrent Nature Biotechnology article describes the development of a mouse...accessible before are now accessible using this technology.” GeCKO library for genomic screens Using lentivirus... Castillo Velasco-Herrera M, Yusa K. Nature Biotechnology. 2013 Dec 23. (Article). Read more about CRISPRs...

...limitations by using gas vesicle reporters that are detectable by an inexpensive and widely available imaging...fluorescent or bioluminescent reporters usually only detects the signal of the reporter and fail to pinpoint...each ARG’s distinct ultrasound fingerprint can be detected. Bourdeau et al calls this method acoustic collapse... in relation to internal organs. The limit of detection for arg1 expressing ECN is 109 cells/ml, which...

...material for a ChIP-seq experiment can be a major technical hurdle.   ChIP-seq requires  cross-linking during...improvement and simplification of the original technique, specifically in terms of initial steps of cell...ChIP-seq, especially in situations when it is technically challenging to collect large numbers of cells...the CUT&RUN method, it seems likely that this technique will quickly become a standard method alongside...

...Snapgene generated maps provide improved feature detection with an easy to use interface. We regularly use... linear or circular sequence. Next, we use the “Detect Common Features” function to find and annotate ...s “Import primers from a list” function, we can detect all of the primers we have designed that will anneal...plasmid maps and sequences use the same feature detection software as the desktop version of Snapgene. When...

...sensitive detection of the plant hormones strigolactones By Fabienne Furt Current methods used to detect strigolactones...CRISPR-based editing in organoid Biosensors for detecting strigolactones The CRISPR Corner New ready-to-...intensity, it provides a direct and quantitative detection of SLs. The authors also showed that the receptor-based...

...purification tools (and can have secondary functions as detection aids.) When a single protein is expressed, with...from solution. MBP antibodies are available for detection and/or purification of MBP-tagged proteins. The...alternative to amylose resin and also provides a visual detection mechanism via applications such as western blot...of interest, especially if downstream antibody detection is desired… and yes, we have an antibody for that...

... be your critical experiments and why? What techniques and skills do you need to use to reach these ...address a topic in a mechanistic way using the technology available.” Choosing a research project “entails...entails good judgement with the current technology to identify a problem, asking what approach will I take... importance of combining “new discoveries or technologies increase chances that your project will be fresh...

...no post-PCR procedures. Sounds easy right? The technique relies on the basic ability of complementary basepairs...called TOPO-TA) cloning; however, the TOPO cloning technique has also be adapted for blunt end cloning. So ... DNA ends to facilitate replication. The TOPO technique specifically uses Vaccinia virus-isolated topoisomerase...Protocols Catch Up On Your Basic Molecular Biology Techniques Watch Our Protocol Videos ...

...AAV viral preps to easily find and share useful technical data. These data focus on experimental design ... These questions are focused on injection and detection methods. For example, you can provide location...you wait until performing the experiment? What detection methods did you use for expression of AAV cargo...Considerations For Precise Neuronal Targeting Using AAV Technologies Using AAV for neuronal tracing Resources on...

...context of IHC, but the problem can impact other techniques like Western blots or immunocytochemistry (ICC... described here may be as effective for other techniques, but it is important to be aware that the issue...Flexible Mouse-On-Mouse Immunohistochemical Staining Technique Adaptable to Biotin-Free Reagents, Immunofluorescence..., Jacob DA (1990) A novel immunohistochemical technique for demonstration of specific binding of human...

...the chance to win some of the latest tools and technologies in life sciences research. The rare disease ...countries. “Over $600,000 worth of cutting-edge technologies were awarded to study 31 rare diseases,” said...ll combine induced pluripotent stem cell (iPS) technology and CRISPR-Cas9 constructs from Addgene to gene-edit...those mutations, they now plan to use CRISPR-Cas9 technology to create cell lines and/or mouse models carrying...

...fluorescent ligands to the loci of interest. Dual color detection In order to label two loci simultaneously, Ma ...CRISPRainbow) to 3-fold with CRISPR-Sirius. Triple color detection With a third sgRNA containing alternating MS2 ...target site. Of these, 26 of the 46 loci gave a detectable signal in human U2OS cells. Based upon this, ...engineered sgRNAs using CRISPRainbow.” Nature Biotechnology 34 (2016): 528-530. PubMed PMID: 27088723 PubMed...

...Positive vs Gram Negative. Technology Networks (2019) https://www.technologynetworks.com/immunology/articles/...