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Showing: 201 - 250 of 954 results
  1. New CRISPR Tools: Cas7-11 and PASTE

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    Blog Post
    ...impact. In molecular and cellular biology, this happens through the development of tools that exploit and...observations, Koonin and Makarova found that Cas7-11 appeared to have derived from the more complex type III-A...experimental design and its potential for clinical applications. Specifically, efficacy rates of Cas7-11 were...patient's specific mutations. With a drag and drop approach, however, the specific mutations wouldn’t matter... the entire gene, allowing a one-size-fits-all approach to gene replacement therapy for most patients....PASTE). Finding the right integrase for this new approach proved a challenge. The old standby of CRE/flp...place, while increasing the number of potential applications. But they weren’t done engineering quite yet...
  2. Antibodies 101: Immunoprecipitation

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    Blog Post
    ...control antibody. Alternative applications Oftentimes, an IP-appropriate antibody simply isn’t available...specificity of anti-epitope antibodies make this approach appealing, but the disadvantages must be carefully ...measure the target. However, the two different approaches allow for optimized strategies for differing ...target protein is then eluted. This method is appropriate for most situations.  A slightly different method...Protein A and Protein G are not suitable for all applications, such as protein isolation from serum. In this...capture antibody interaction and/or the downstream application. Typically, if the target protein will be detected...
  3. Microbes: Look, Listen, and Tell

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    Blog Post
    ...cellular imaging with ultrasound. In his research he applies naturally occurring bacterial “molecular buoyancy...biomolecule-based dyes. This technology is widely applicable as bacterial expression of dyes allows dye visualization...acoustic reporter genes’ on our blog. Creative approaches to presenting the microbial world After learning...the lab, I was excited to learn about creative approaches to visualize bacteria in an educational setting...setting. The Symposium on ‘Creative Approaches to Presenting the Microbial World’ showcased complex connections...connections and ideas in various visualized approaches from hand-drawn comics (Dr. Sanja Saftic) to educational...makes scientific topics both interesting and approachable!   The grand finale of visual impressions was...
  4. Let There Be LITE Plasmids

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    Blog Post
    ...effectors (TALEs) as anchors for genome engineering applications was still very new. Now, Konermann and her colleagues...just how flexible their LITE system could be by applying it to altering gene expression via epigenetics...light retraction. Finally, because light can be applied non-invasively, its use is not restricted to cultured...optogenetic tools." In total, the researchers applied the LITE system to 28 different genes to establish...cellular processes." The LITE system can now be applied to unraveling the genetic and epigenetic players...disease states. While LITE plasmids have clear application to anyone with an interest in neuroscience, their...elegans and in Plasmodium. For those who do want to apply LITEs for the transformation of neurons, she offers...
  5. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

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    Blog Post
    ...experimental approach you choose, it’s essential to consider the technical limitations of each approach and to...affect the phenotype. Viral vectors and genetic approaches are continuously refined and developed so there.../loxP or Flp/FRT, with AAV vectors are common approaches to control transgene expression by restricting...post-translational regulatory elements. Examples of these approaches are given in Tables 3-7. 3. Making use of AAV...populations using viral strategies Combining different approaches can increase the specificity of cell targeting...experimental design and data. With these different approaches to open science, we aim to accelerate the rate...
  6. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

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    Blog Post
    ...Polyethylene Glycol (PEG), although the application of this approach is limited to protoplast cells (plant...current CRISPR research focuses on its potential applications for human medicine (Waltz, 2016), the potential...desirable traits using traditional plant breeding approaches, these techniques are cumbersome, often requiring...selected the correct CRISPR components for your application, it is time to deliver these components to your...find many plasmids for a wide variety of CRISPR applications in plants available through Addgene. In addition...N (2016) The CRISPR/Cas Genome-Editing Tool: Application in Improvement of Crops. Front Plant Sci 7: ....10.1073/pnas.1818290116 Zhu H, Li C, Gao C (2020) Applications of CRISPR–Cas in agriculture and plant biotechnology...
  7. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

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    Blog Post
    ...in the blood. But the virus doesn’t completely disappear; it just hides in latently infected cells. To ...of the integrated HIV-1 genome. For a targeted approach, Zhang et al. turned to the CRISPR/Cas9 Synergistic... essential to HIV-1 replication. However, this approach could permit some viral protein production, so...animal models, and it’s difficult to tell if one approach might be more successful. “Shock and kill” has...HIV-1 to deplete the viral reservoir, and this approach doesn’t require a functional Cas9 nuclease, so... reservoir spans multiple organ systems. Both approaches may prove more successful early in the course...for either “shock and kill” or viral excision approaches, and each of these gRNAs will need to be validated...
  8. Viral Vectors 101: Systemic Capsids

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    Blog Post
    ...suitability of a systemic capsid for an individual application is dependent on a number of factors, including...test your chosen capsid before using it in an application.     Figure 1: AAV toolkit for targeted...doses are recommended for all systemic capsid applications. For example, AAV-PHP.eB is typically administered...capsids or transgenes. Sensory neurons and liver appear especially susceptible to off-target transgene ...experimentally determine the best titer for your application.   Pro tip!Want to know if a capsid is a good...always, validate the capsid for your specific application and animal model.  Other model organisms Tropism...astrocytes or oligodendrocytes when paired with appropriate cell type-specific promoters or Cre lines. Spatial...
  9. Antibodies 101: Epitope Tags

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    Blog Post
    ...experienced. Finding an antibody that works for your application, specifically detects your protein, is species...and have been widely used across species and applications. In this blog, we will review the most common...not exist, or they don’t exist for a specific application, or they exist but are unreliable for a variety...almost certainly already been optimized for your application. Depending on the tag, directly imaging or tracking...our affinity purification tag blog!).   Applications of epitope tags   The tags A number of...with moderate to high success rates across all applications. You can find FLAG antibodies on Addgene’s catalog... biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol. (2003) 60:523-533 DOI: 10.1007...
  10. Hot Plasmids: FRET-Based Biosensors

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    Blog Post
    ...out that the approach he and his colleagues took to optimize sensitivities can now be applied to dozens ...Because they are chemical compounds, they have to be applied or injected, and they don’t always end up where...lymphocytes, too, making them a versatile tool for application in both neuroscience and immunology. Don’t be...FRET-based sensors. For example, his team is now applying it to improve upon an indicator they developed... calcium versus zinc should require the simple swapping of binding domains, in practice optimizing the... large dynamic range,” he says. Merkx recently applied this “trick” to the development of additional color...
  11. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

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    Blog Post
    ...reproducible if its findings can be predictably applied to future work. Importantly, no one should expect...studies, as reported by drug companies attempting to apply the findings from these studies, are quite low (...low rates of reproducibility. Failed attempts to apply irreproducible results to future work cost both ...solutions based on what I’ve learned. I hope you can apply some of these solutions to your own research and...ridiculous to expect that one should be able to apply someone else's new and incredibly useful technique...information about fluorescent proteins so you can best apply these technologies to your own research needs. Other...evidence shows otherwise. Others will go on to try to apply these findings in their own work ultimately wasting...
  12. Technique: Probe Phage Genomes for Host Binding Proteins

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    Blog Post
    ...interesting, much of what a phage faces as it approaches a host cell surface are complex polysaccharide...involves digesting phage genomic DNA into 1-3 kb (approximately gene-sized) fragments so that these fragments...competent E. coli and analyze transformants for appropriately sized inserts. We consider a library worth using...-side up onto a fresh LB plate containing an appropriate inducing agent (0.4 mM IPTG is a good start if...organism and incubate overnight according to appropriate cell growth conditions. Note: if using fast-growing...Franklin L., et al. "Revisiting phage therapy: new applications for old resources." Trends in microbiology 23.4...Mirzaei, and Anders S. Nilsson. "Adapting drug approval pathways for bacteriophage-based therapeutics....
  13. Finding Your Perfect Job After University

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    Blog Post
    ...received many applications, employers were requesting that only scientists with an MSc or higher apply. It was...scientific research would be very competitive, and so I applied for a summer role as a Research Scientist at the... edge of London to gain relevant experience. I applied for this summer position as I felt I needed more...I had to reassess what I wanted to do and so I applied for a role in Industry. I wanted to get "a real...university to study a Masters of Science I decided to apply to study a Master of Science (MSc) in Human Molecular...virology classes in California and enjoyed them, so I applied for a role at a commercial virology testing company...enjoy helping people and decided to compromise and applied anyway. I later found out the job was with Addgene...
  14. Using CRISPR/Cas9 to Edit Disease Out of the Genome

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    Blog Post
    ...the technology in different labs for different applications.” Li’s team noted that CRISPR/Cas9 has been ...disease. Meanwhile, Hans Clevers and colleagues applied CRISPR to disease correction in adult stem cells...to be perfected until the efficiency of repair approaches 100 percent. “We need to be sure it's accurate...repair,” Doudna said. “And it can't be too hard to apply. These papers suggest it might work well enough ...with size limitations. In addition, non-viral applications are also possible through the use of nanoparticles....  “CRISPR Genome Editing Technology and its Application in Genetic Diseases: A Review.” Current pharmaceutical...4. Uddin et al. 2020.  “CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future...
  15. Trading Spaces - Visiting Scientist Edition

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    Blog Post
    ...out some feelers, apply to programs, and apply for travel grants. Good luck and happy travels! Resources...workdays will be carried out until the end of the appointment. The visiting scientist could be a graduate student...visiting positions within defined labs which you can apply to. These structured types of placements often have...are your options if neither of these situations apply to you? Once you’ve identified one or more labs ...mentor can help get the ball rolling! If you are applying to a structured program, they likely have guidelines...
  16. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

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    Blog Post
    ...and most patients still don’t benefit from this approach. One central bottleneck is that most recurrently...computational algorithms, such as SIFT or PolyPhen to approach this challenge, using attributes of cancer variants... would be extremely helpful as a complementary approach. Also, in principle, these experiments could find...sequencing studies (as well as reference cDNA clones mapping to well-annotated cellular pathways). It wasn’t... designed to complement the one assay per gene approach that has been the historical standard. Specifically...team have recently demonstrated the power of this approach: Phenotypic Characterization of a Comprehensive...technical feasibility and proof-of-concept. If such approaches are proven to improve future clinical decisions...
  17. Hot Plasmids - May 2022

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    Blog Post
    ...Proximity-dependent labeling has become a powerful approach for proteomic mapping in both cell culture and whole organisms...organisms (see our 2019 overview). Until recently, the approach has been hampered by the need to tag each protein...provides researchers better flexibility and makes the approach more accessible, as it reduces the need to generate...optimized with zebrafish in mind, it can easily be applied to other systems.    Find BLITZ Plasmids here! ...Targeted Evolution method (M-CREATE), in which one applies both positive and negative selective pressures ...clonal multiplexing of cells in live imaging application. Each visual barcode consists of a fluorescent...
  18. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    Blog Post
    ...RNAi) is a conserved eukaryotic process where approximately 20-30 nucleotides of double-stranded RNA (dsRNA...silencing can be easily induced by exogenous application of long or short dsRNA. However, it is important...have benefited from the ease of use in dsRNA applications. Figure 1: Schematic of an RNAi-inducing... two. Consequently, I will focus on different approaches and considerations present at the first stage...long-term RNAi phenotype. Long hairpin RNA The easiest approach to induce RNAi involves the expression of long...RNA The greatest limitations of the long hpRNA approach are thought to be off-target effects and variable...your gene(s) of interest and the reporter. This approach greatly enhances experimental throughput, as silencing...
  19. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

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    Blog Post
    ...best for insertions or deletions greater than approximately 20 nucleotides, while ssDNA substrate is best...regions flanking the target site).  The best applications for dsDNA inserts include: large insertions ... used in limited circumstances. An alternative approach is to flank the desired change with 4-5 silent...Uses in Recombineering Substrate Best Suited Application Recombination Frequency Special Considerations...can be modified with any of the below described approaches.    Bacterial strain with integrated defective...requires drug selection.  One drawback to this approach is that it requires the generation of bacteriophage...modification into a different background, if needed. This approach is best suited for modifying plasmids or BACs ...
  20. Human Germline Editing Using CRISPR

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    Blog Post
    ...While repair using endogenous chromosomal DNA is appealing for heterozygous mutations, it will clearly be...consider that already-tested CRISPR alternatives are applicable in many situations. In the case of hypertrophic... need to be much, much lower before clinical applications could be considered. It is important to consider...consider the question of broader applicability of CRISPR germline editing. Nature News mentions that the...between CRISPR repair in embryos and iPS models, it appears that embryo research is the only way to answer ...answer questions of efficiency and applicability. Given moral and ethical concerns surrounding such research...experiments are needed before we can consider clinical applications for germline editing. Although this paper represents...
  21. Transferable Skills Guide: Problem Solving

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    Blog Post
    ...definition of “problem solving skills” is the ability to apply the scientific method to problems in a variety of...strong problem solving skills is just a matter of applying that framework to the myriad challenges we encounter...always found that taking a conscious or active approach can be hugely helpful not only for developing ...ask them to explain their logic. Or you can try applying the scientific method to everyday problems in ...about technical know-how and more about how you approach new challenges.     Know your strengths! Research...to practice problem solving skills and learn to apply them flexibly in a range of contexts. As such, scientists...remember that you have these skills when you start applying and interviewing for new positions. Highlighting...
  22. New Tool for Lineage Tracing: The ClonTracer Library

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    Blog Post
    ... the library can be used to identify appropriate, non-overlapping targets for drug development - such ...industry research. When she came to Novartis, she was happily surprised that “Novartis was actually really research...were so many interesting questions that we could apply [it to].” Using ClonTracer to study drug resistance...using NGS because the error rate of current NGS approaches allows for a detection limit of roughly 0.1%....populations can be eradicated up-front quickly. This approach is in contrast to the current treatment paradigm...Novartis Oncology and is particularly interested in applying what we have learned about tumor heterogeneity...
  23. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ...assembly process designed for a quick, plug-and-play approach to cloning CRISPR vectors (McGee et al., 2024)... plug-and-play. Because of its modular cloning approach, it’s easy to switch out reporters, Cas proteins...overview and timeline of the Golden Gate (GG) cloning approach. Individual modules and a sample ligation are ...validated and were chosen because of their broad applicability in different model systems. What’s the error...  The Fragmid website While the modular cloning approach is key to the Fragmid system, the Fragmid website...sites needed for the Fragmid cuts, clone in the appropriate overhang for your module type, and voilà! You...great plasmids for your next CRISPR experiment. Happy cloning! References and Resources References McGee...
  24. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

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    Blog Post
    ...insertions or deletions in the DNA sequence. This same approach is effective to target short ncRNAs, such as microRNAs...adjacent genes, especially those regulated by or overlapping with the lncRNA (Basak and Nithin 2015, Horlbeck...A schematic illustration of CRISPR/Cas9-based approaches to study ncRNAs. (A) Transcription start site...it comes to targeting precision and practical application. If the CRISPR/Cas9 system is not effective for...to enhance or upregulate ncRNA expression. This approach is useful for lncRNAs because they regulate nearby...lncRNAs with high specificity, particularly in applications requiring smaller delivery systems (Zibitt et...science.1225829  Hazan J, Bester AC. CRISPR-Based Approaches for the High-Throughput Characterization of Long...
  25. The Materials Science of Optogenetics Experiments

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    Blog Post
    ...patch cable must be the appropriate length for your specific behavioral apparatus or test but not so excessively...the initial laser output. The same considerations apply to the polishing and preparation of the patch cable...experiment started, you will need a laser of the appropriate wavelength to stimulate your opsin. Each opsin...to optogenetics users and adjusts the lasers appropriately (tuned for optogenetics). These lasers are relatively... under certain contexts/locations in the test apparatus and thus individual laser stimulation is essential...Injectable, cellular-scale optoelectronics with applications for wireless optogenetics. Science. 2013;340...
  26. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...potential applications of CRISPR/Cas9 in genome-engineering. One particularly attractive application involves...cargo RNA to the appropriate DNA locus. The sgRNA/RNA fusions retained their appropriate functions affecting...cleave the target DNA. The inducible split cas9 approach can also be used for activation of specific genes...necessary light-activated protein domains to the appropriate locus has been cumbersome. To overcome many of... system widely accessible for many potential applications. Polstein LR & Gersbach CA, Nat Chem Biol ...amenable to both high and low throughput screening applications. The system can be readily modified to use different...the stop codon. Thus a non-translated mRNA will appear yellow, but a translated mRNA will be red. This...
  27. Using AAV for Neuronal Tracing

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    Blog Post
    ...iontophoresis (applying small electric currents via inserted electrodes) may be applied, but these methods...intracellularly spread away from the site of application and aid in visualization of the morphology of...the hippocampal region CA2 allowed for detailed mapping of the neuronal circuitry involved in learning ...Mouse Brain Connectivity Atlas website. A similar approach was used by researchers from the Janelia Campus...activity-dependent neuronal tracing. Apart from mapping neuronal networks, improved AAV vectors could also...into muscle) as opposed to direct intracerebral application, which is more invasive and has a much higher...Cre-dependent, anterograde transsynaptic viral tracer for mapping output pathways of genetically marked neurons. ...
  28. Plasmids 101: 5 factors to help you choose the right cloning method

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    Blog Post
    ...processes happen in a matter of minutes. Cost: Funding is usually limited - what cloning approach should...afraid to explore multiple approaches to find out which one works best for you. Happy cloning (and be sure ... of the post! Speed: Your grant deadline is approaching or your advisor is demanding results. If you’...however, or simply looking for a more streamlined approach, you might want to give Gateway a try. This method...LR Clonase enzyme mixes. After generating the appropriate inserts by PCR, there is no need for restriction...
  29. Antibodies 101: Producing Recombinant Antibodies

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    Blog Post
    ...production system may depend on what downstream applications you are planning, cost, and the amount of protein... for this. Chromatography columns containing appropriate immobilized ligands are incubated with cell culture...Quantification Once you’ve collected your antibody in the appropriate buffer, it’s time to quantify it. There are several...accurate concentration to give your downstream applications the best chance at being successful.  Using ...they bind their antigens specifically. Possible applications for validation include western blot, immunohistochemistry...some antibodies are better suited for certain applications. An antibody that performs well in immunohistochemistry...use! You can learn more about common antibody applications in our Antibodies 101 blog series.  Resources...
  30. Course-Based Undergraduate Research Experiences (CUREs)

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    Blog Post
    ...to take a different approach. His own most impactful learning experiences happened in the lab, and he ...experiences afforded to those fortunate enough to apprentice with top scientists to larger groups of trainees...notes the importance of acknowledging student apprehensions about “failure” (protocols not working, hypotheses...while reminding participants of the value of this approach and encouraging them to engage in the problem-...in setting up a CURE, especially if it’s a new approach within their department. So in 2016, Provost partnered...research-based laboratory – a semester-long research approach using malate dehydrogenase as a research model...
  31. Lentiviral Vector Uses and Overview

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    Blog Post
    ...many CRISPR applications, and they’ve also had success in clinical gene therapy applications. Read on to...to learn more about the current (and future) applications of lentiviral vectors! Lentiviral vector history...barcoding libraries have been used for complex applications like lineage tracing in heterogeneous tumors...developed integrase-deficient lentiviral vectors for applications that require the large genetic capacity of a...treatment of a molecular disorder: gene therapy approaches to sickle cell disease. Hoban MD, Orkin SH, Bauer...Integrase-defective lentiviral vectors: progress and applications. Banasik MB, McCray PB Jr. Gene Ther. 2010 Feb...
  32. CRISPR 101: Targeting RNA with Cas13a (C2c2)

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    Blog Post
    ...use, functional in most species, and has many applications. That said, SpCas9 is not the only game in town... makes this molecule unique, and the various applications of Cas13a. The origins of Cas13a: An RNA cleaving...identity to the spacer sequence and contain the appropriate PAM sequence. The non-specific cleavage is thought...discussed below. Subscribe to CRISPR Blog Posts Applications of Cas13a   Figure 1: Using Cas13a as...of nucleotides.   What are the potential applications of Cas13a given what we know about its structure...CRISPR toolbox and will continue to expand the applications of CRISPR. Since this post was originally published...development seen in the field and as always, we appreciate the opportunity to share these invaluable reagents...
  33. To Codon Optimize or Not: That is the Question

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    Blog Post
    ...may be found at very different levels (Mauro and Chappell, 2014). Due to this variation in tRNA expression...is codon optimization? Codon optimization is an approach in gene engineering to improve gene expression...sites and even affect protein function (Mauro and Chappell, 2014). Different rates of translation by different...and in biotechnology and therapeutics (Mauro and Chappell 2014).  Ultimately, it is important to take a ...//doi.org/10.1186/s12859-017-1793-7 Dresios J, Chappell SA, Zhou W, Mauro VP (2005) An mRNA-rRNA base-...https://doi.org/10.1101/gr.139758.112 Mauro VP, Chappell SA (2014) A critical analysis of codon optimization...synonymous codon usage bias, and its potential applications. Nucl Acids Res 15:1281–1295 . https://doi.org...
  34. Career Insights: Technical Support Specialist

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    Blog Post
    ...study for late career scientists to become more appealing on the job market (e.g. a postgraduate degree ...so much better. Driven by that, I successfully applied and worked as a technical support specialist for...expertise in technical support positions. When applying for technical support positions, you should try...company. To increase your chances at successfully applying for a position, you should familiarize yourself...instances, variations of the product’s specific application instructions or “optimized steps” can be linked...customers may tell all sorts of details about the application, just not the ones that matter. Therefore, if...different backgrounds and realizing that the same approach does not work for everyone. The benefit is that...
  35. Healthcare Consulting: A Door to the Business of Life Sciences

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    Blog Post
    ...requiring people who are willing to quickly learn, and apply their knowledge to a variety of problems. In this...chemotherapies, or developing innovative mobile phone applications and services that help diabetic patients manage...Personally, I have greatly benefited from and appreciated the training I’ve received in concisely and accurately...literature, and critical analyses skills were applicable to my cases. I can directly use my understanding...core of the suite of benefits a given therapeutic approach brings to patients, doctors, and insurance providers...the gap between the science and the commercial applications of a particular therapy. With your scientific... research and bench science, with a newfound appreciation for how technologies get developed and commercialized...
  36. The Addgene Antibody Data Hub

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    Blog Post
    ...very little data on which might work in your application. It can be quite difficult to decide on which...be searched and sorted by gene/target, name, application, pass/fail rating, sample species, antibody species... how the antibody performed in the submitted application, along with a brief description of the experiment... section will vary slightly depending on the application — flow cytometry, western blot, immunohistochemistry...data showing how the antibody worked in your application. The submission process is easy, with simple,...simple, descriptive fields, drop-down boxes when appropriate, and can be saved at any point throughout the ...
  37. Quick Guide to Near-Infrared Fluorescent Proteins

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    Blog Post
    ...robotic multidimensional directed evolution approach applied to fluorescent voltage reporters. Nat Chem...information on NIR FPs, their properties and applications. At the end, there is a Table that summarizes...phytochrome (BphPs) photoreceptors have higher apparent cellular brightness than previously reported widely...their enhanced versions named emiRFPs in your applications.  There is a whole set of spectrally distinct... about the characteristics, performance, and applications of near-infrared calcium indicators in this ...of Medicine. Her expertise is development and applications of fluorescent proteins, genetically encoded...
  38. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

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    Blog Post
    ... Brainbow 3.2. Brainbow labeling has also been applied to cell marking and lineage tracing in other systems...Between Brainbow Systems   Brainbow variants and applications beyond the mouse brain In addition to improving... Currently, Brainbow techniques are also being applied to study model organisms such as Drosophila and...zebrafish. Brainbow in Drosophila has aided in the mapping of neural circuits, such as connections between...Brainbow’s colorful labeling has decreased the mapping time for a given section of brain by at least an...brain and other biological systems. Interested in applying Brainbow to your research? Check out Brainbow ... Nov 1;450(7166):56-62. PubMed. A technicolour approach to the connectome. Lichtman JW, Livet J, Sanes...
  39. Optogenetics + CRISPR, Using Light to Control Genome Editing

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    Blog Post
    ...improvements to CRISPR technology since its initial applications for genome engineering in 2012.  Many of these...particular interest since blue light has limited applications in vivo. UV and blue light is readily absorbed...allowing DNA binding and cleavage. This technique was applied to create photoswitchable spCas9, dCas9, and S....saCas9) proteins, showing the versatility of this approach. This study generated the first optically active..., allowing the release of Cas9. A noteworthy application of the CASANOVA system was its use in light-mediated...Deiters’ lab, on the other hand, took a different approach. These researchers used a genetically encoded ...Photoactivatable moiety Cas9 variatn CRISPR protein application Split Systems / Light-induced Dimerization ...
  40. Deep Dive: qPCR

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    Blog Post
    ...understanding and technical proficiency with common applications - now let’s dive right in! You know qPCR is...as it solves, so understanding and using the application correctly is imperative. Picking your chemistry...space. For many cell lines and tissue types, appropriate reference genes (and supporting data) are often...2002), though there are cases in which it is an appropriate choice (Panina, 2018). Look for studies specifically...Vandesompele, 2002) will help you select the appropriate reference genes - and it is well worth your time... more, take a look at their 551 page e-book on applying MIQE guidelines! With these kinds of resources...melting profiles of PCR products in a rich web application, Bioinformatics, Volume 27, Issue 7, 1 April ...
  41. Bringing Sustainable Practices to the Lab: Innovations

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    Blog Post
    ...New ideas and approaches are encouraged in science. It isn’t surprising that new ideas are also popping...lab lives. Read on to learn about some creative approaches to lab sustainability, and find ways to further...but what about all the manufacturing steps that happen before you even receive the product? How can we...tips. This is part of a cultural change in how we approach waste that needs to be supported at all levels... facility, where it’s sanitized through an EPA-approved process and no longer considered hazardous. Then...industrial and residential markets. Rethinking our approach to lab waste is something we all need to do. Remember...
  42. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ...Early CRISPR applications were often limited by the low editing efficiency of homology-directed repair...BE3, nicks the unmodified DNA strand so that it appears “newly synthesized” to the cell. Thus, the cell...reduce the undesired C→G or C→A conversions that can happen with BE3s. These byproducts likely resulted from...making them a promising tool for cell therapy applications. Dual base editors What about combining the ...Grünewald et al., 2020). Another lab took a similar approach. Dali Li’s lab’s A&C-BEmax consists of a fusion...activity compared to ABEmax. In an alternative approach, engineering and directed evolution of the TadA...editors with increased activity and therapeutic application. Nature Biotechnology, 38(7), 892–900. https:...
  43. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ...of the protein of interest. An electric current applied to the gel causes the negatively charged proteins...with blotting paper and an electric current is applied causing proteins to migrate out of the gel and ...sandwiches.   Figure 2: The western blot dry apparatus includes a stack of filter paper, a copper anode...search, first pay attention to the validated applications listed on the vendor’s website and choose an...western and vice versa.  In addition to validated applications, pay close attention to the controls listed ...choose antibodies that are validated for your application and test the antibody in your specific system...to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences...
  44. Recombinase-based State Machines Enable Order-dependent Logic in vivo

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    Blog Post
    ...attB-attP pairs from different recombinases are overlapping or nested on a register, then the operation of...more population-wide (as opposed to single-cell) approach is to interrogate state with qPCR. The re-arrangement...performed on one register at a time. If a particular application makes it difficult to isolate single registers...state read-out method may not be suitable for applications that require minimal cellular perturbation. ...contains the register. Depending on the desired application, the inducible promoters we used to drive recombinase...program to find the one that best suits your application. How efficiently do RSMs work? We tested a two-input...anticipate that, depending on the context and application, some inducible recombinase systems may work ...
  45. Year of the Camelid: Antibody Style

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    Blog Post
    ...diagnostic applications. For example, fluorescently and radioactively labeled sdAbs have become appealing candidates...to sdAbs becoming valuable tools for diverse applications. For example, RANbodies can be used as an alternative... as well. The first sdAb based therapeutic was approved in 2018, with many others in clinical trials (...maintaining a llama facility is out of reach and unappealing to many labs), the field is moving towards using...inspired this versatile family of tools, the applications they have already enabled, and the discoveries...
  46. We're Updating Our Hold for Publication Policy

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    Blog Post
    ... complete necessary quality control and legal approvals so the plasmids are ready for distribution as ...this Hold for Publication policy seriously and appreciate the trust you’ve put in us when you choose this...worry, we’ll let you know before these releases happen (and congratulate you on your paper!). And as always...your deposit or plasmid ID numbers and we’ll be happy to update the plasmids to add the citation and release...hope somebody will find the plasmids useful, we’re happy to distribute unpublished materials. What about ...needs, so please let us know what you think. We appreciate your feedback. If you have questions about depositing...
  47. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    ... be used as a fusion tag for a wide range of applications from traditional imaging to FRET and stochastic...transforming E. coli with a gene library of approximately 484 variants, including some potential tetramer-breaking...Florida State University, they used a mutagenesis approach and found that specific amino acid changes favored...violet range. For example, mWasabi can be used with Sapphire (excitation maximum 399 nm) because it does not...fluorescent protein: structural characterization and applications in fluorescence imaging." Biochemical Journal...identification of the molecular determinants of color and applications in fluorescence imaging." BMC biology 6.1 (2008...
  48. Important Considerations When Using AAVs

    Type
    Blog Post
    ...possible tools, lentiviruses for example, are not approved for use in vivo use at the University of Manitoba... be mostly on the brain, but many of these tips apply to all tissues. Subscribe to Viral Vector Blog Posts... infect) can influence an AAV’s value greatly. Applying this mantra to AAV selection, there are three ...of AAVs? 1) Combined functional and anatomical mapping of circuits In our lab, we use AAV-DREADD complexes...only can we compare these methods in specific applications, we may also be able to use these comparisons... on overall cell function. AAVs are the most appropriate vehicle for this type of research, as they are...efficiency, and safety profiles should strengthen its appeal for the scientific and medical communities. Many...
  49. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ... Many thanks to our guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at theJanelia... have been completed. Therefore, to be ready to apply for a Group Leader position, key first author manuscripts... project. Furthermore, Group Leader position applications require 3 letters of reference- the graduate...informal recommendations, more positive grant application and manuscript reviews, etc. You'll also meet... fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance runner, gardener...
  50. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    ...information they’ve absorbed if they don’t get a chance apply it in an interactive setting. The problem is…how... as I try being compelling, interactive, and application-driven to boost curiosity, I have never experienced...they would use their logic and strategy skills to apply those concepts and understand how therapeutic strategies... BUILD YOUR PLASMID is a game where you need to apply basic molecular biology, cell biology, microbiology...best strategy and propose a final gene therapy approach for the disease to be openly discussed with the...opposing team. In the end, the students had fun while applying most of the concepts required by the course and...
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