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We narrowed to 812 results for: APP

Showing: 101 - 150 of 812 results

  1. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    Published
    April 14, 2022, 1:15 p.m.
    ...allow you to be flexible in your approach to affinity reagent applications. Whatever affinity reagents you...re used in a number of research and clinical applications. Most of the time, when someone speaks about...deeper penetration into a tissue, useful for applications like IHC, or be able to access an epitope folded...that would size-exclude an antibody. In imaging applications, smaller affinity reagents means higher resolution...antibody’s ‘Y’. They are often used in clinical applications. Fab fragments can be created in lab in situations...Shahied, 2004). They are often used for clinical applications, including imaging.  Diabodies are ~60 kDa proteins...antibodies are still an excellent option for many applications, and frankly, if ain't broke, don't fix it!  ...

  2. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    Published
    March 6, 2018, 1:49 p.m.
    ... chain reaction (LCR)-overview and applications." PCR Methods Appl 3.4 (1994): S51-64. PubMed PMID: 8173509...or DNA base pair mismatches (8-11). For some applications, there cannot be any end-joining activity, and...regions. Why use a high fidelity ligase? Numerous applications have been developed to take advantage of the...Taq and other thermostable DNA ligases. These applications detect specific nucleotide sequences with high...Ligase are not replacements for T4 DNA Ligase in applications such as routine cloning or DNA library preparation...Temperature Calculator can be used to estimate an appropriate incubation temperature when using NEB thermostable...Christopher D. Lima. "The polynucleotide ligase and RNA capping enzyme superfamily of covalent nucleotidyltransferases...

  3. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    Published
    Oct. 4, 2016, 2:30 p.m.
    ...sequencing is laborious, time consuming and cannot be applied to heterogenous populations (Bell et al. 2014).... on genome sequencing. Used in concert, these approaches can provide the researcher with a reasonable ...variable) Sequence-level data Low-throughput, inappropriate for heterogenous populations, low sensitivity...(Fu et al. 2013; Pattanayak et al. 2013). This approach misses larger mismatches (e.g. six nucleotides...Genotyping Options Technique Detection Limit Application Advantages Disadvantages GUIDE-Seq 0.1% Cell-based... may be to integrate both biased and unbiased approaches. As suggested in a review by Tycko et al., 2016...a highly accurate manner. Using both of these approaches may not be necessary for every CRISPR experiment...

  4. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ...gRNA targeting approach is then used to direct the complex to a specific locus. This approach provides a ...tools available for bacteria and some of the applications for which they’ve been used. The beginnings ... E. coli. Furthermore, to demonstrate wider applicability they demonstrated success of Cas9 mediated repression...method in E. coli, Jesse Zalatan’s lab used an approach to identify known proteins involved in the activation...scenario allows for several potential CRISPR applications. First, CRISPR can be used to target locations...resolve them, and the cells die.  This could be applied to diseases where the presence of just a single...polymorphisms, including antibiotic resistance genes. This approach successfully targeted specific organisms when ...

  5. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    Published
    Aug. 30, 2016, 2:30 p.m.
    ...need for low-cost, field-based diagnostics, our approach could make an impact. Our motivation was underscored...involved in the effort. This inclusive and open approach by all involved definitely contributed to our ...). Using toehold switch-based RNA sensors, our approach uses portions of the pathogen’s genome as unique...portable quantification of diagnostic results.  Our approach now detects samples at clinically relevant concentrations...longer-term aim is to advance our programmable approach to other critical needs and to further develop...focused on developing cell-free synthetic biology applications for human health and the environment.     Alexander...nanomaterials science, and self-assembly for applications in health, energy, and sustainability.   References...

  6. Transferable Skills: Negotiation

    Type
    Blog Post
    Published
    March 3, 2022, 2 p.m.
    ...mainly do over the price of a used car. Later, as I applied for jobs, I knew I should negotiate my salary after...vacation (New York City or Hawaii?). So how can you approach improving this transferable skill? Negotiation...one of us wins, the other loses. Although this approach to negotiation is the most straightforward, it...it can mean that everyone leaves disappointed (we’ve all won an argument but felt bad afterwards). Or ...there’s no agreement at all and everyone leaves unhappy.   Negotiation as a creative problem-solving process...process Although at first glance it may appear that there has to be a winner and a loser when negotiating...negotiating, what if we instead approached it as a creative problem-solving exercise? Asking questions like ...

  7. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...review how these indicators work, their general applications, and why viruses are essential to their use ...this later). Chemigenetic sensors take a hybrid approach and utilize both a genetic component as well as...sensing to signal output slightly slower than other approaches. Functionally, the voltage sensitivity of the...improvement. These sensors still have many useful applications, though, and if you’re in the market for a VSD...tissues depths (1 mm), important for certain applications such as in vivo tissue imaging.   Figure...your cell type of interest. For in vivo imaging applications, the ability to direct your voltage sensor selectively... as biological tools, our blog has you covered. Happy voltage sensing!   Figure 3: Components ...

  8. Antibodies 101: Flow Compensation

    Type
    Blog Post
    Published
    May 16, 2024, 1:15 p.m.
    ... necessary to apply a compensation value between the two channels (Figure 1A). Applying compensation means...your single-color controls as a starting point. Applying compensation to your experimental samples In order... in the plot. A compensation value of five was applied to APC-Cy7 against APC to correct for this bleed...compensation value of 3.7 for APC into APC-Cy7 applied next, which fixed the bleed in this population....each plot.  The general rules of compensation apply to your experimental samples as they did with your...populations and may need to be “comped” first before you apply compensation values to your current channels. A ...plot, and the population does not move even after applying very large compensation values (100+), consider...

  9. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    Published
    Jan. 19, 2021, 2:15 p.m.
    ...tool that can be used for a variety of research applications. Within the immune system, antibodies are produced...antibodies to bind proteins is useful for research applications as well because they allow scientists to target...150 kDa, a nanobody is 12-15 kDa.   Research applications for antibodies You can use antibodies for experiments...method of detection. You’ll also want to use the appropriate controls for your experiment to account for potential...Let’s dive in and take a look at some of these applications. Western blot Western blots are used to detect...quantitatively if you also load the gel with appropriate controls for normalizing the protein concentration... by binding to cell surface receptors. These applications are typically used for investigating signaling...

  10. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    Published
    March 21, 2024, 1:15 p.m.
    ...emission wavelengths, which are pivotal in their application in biotechnology (Grimm & Lavis, 2022). These... substrates/peptides and antibodies. Example applications of this include coupling to an antibody, whereby...typically bind to. Stability of fluorescence: For applications that involve ranges of pH, and significant lengths...light, is another factor to consider. If your application requires longer periods of exposure, you’ll want...descriptions, or technical manuals. Specific applications and matching dye/ligand choices For effective...Fluorescence Resonance Energy Transfer (FRET) applications, the dyes need to have compatible excitation...more stable fluorescence for super-resolution applications. Other current advancements include work exploring...

  11. Transferable Skills Guide: Identifying Your Transferable Skills

    Type
    Blog Post
    Published
    Jan. 11, 2018, 3:29 p.m.
    ...not-so-technical skills you’ve built over time are only applicable to those positions you have already held. Imagine...have the desired degree, you have mastered the applications they use, you’ve gained experience working with...that you will contribute openness, clarity, approachability, and professionalism to team meetings and projects...identified by another person. Similar reasoning may also apply to other acquired skills.” It is nice when other...second to reflect. For example, say your boss approaches your team with a project she needs one of you... have leadership skills that are just not being tapped at the moment? You can take this time to step up...more of a leader than you thought! When change happens at your organization, when your boss needs someone...

  12. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ...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...

  13. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    Published
    Feb. 17, 2022, 2 p.m.
    ...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...

  14. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    Published
    April 19, 2018, 1:25 p.m.
    ...becoming increasingly important as more complex approaches to scientific questions become popular. There... and don’t be afraid to make something concrete happen. You may have more willing collaborators than you...and meeting people with different mindsets and approaches can truly broaden your mind. When I first began...felt that I had to overcome some hesitation to ‘approach strangers’ and learn how to communicate outside...somewhat awkward in the beginning. But, as so often happens, one gets better with repetition. I learned to ...and started to be fun. I just had to remember to approach the relationship with the perspective of my potential...collaborators in mind. Being at a university allows you to approach people outside the university within the context...

  15. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    Published
    April 11, 2017, 2:30 p.m.
    ...the appropriate ligand, but relying on a lack of fluorescence for readout may render this approach less... Aptamer biosensors are used in a variety of applications, including detection of environmental pollutants... opens aptamers for use in many entirely new applications including aptamer based fluorescent biosensors...less sensitive in most applications. Learn About Fluorescent Biosensors Light-up aptamers (or, fluorescing..., MG proved to be too toxic for many in vivo applications, but this work proved the concept that RNA could...and biochips. Ultimately, there are just some applications that lend themselves to aptamer-based instead...Kyung-Mi, et al. “Aptamers and Their Biological Applications.” Sensors (Basel). 2012; 12(1): 612–631. PubMed...

  16. Microbes: Look, Listen, and Tell

    Type
    Blog Post
    Published
    Aug. 28, 2018, 12:04 p.m.
    ...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...

  17. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ...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...

  18. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    Published
    Nov. 27, 2019, 2:03 p.m.
    ...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...

  19. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    Published
    Oct. 11, 2016, 2:30 p.m.
    ...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...

  20. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    Published
    May 10, 2016, 2:30 p.m.
    ...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...

  21. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ...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...

  22. Antibodies 101: Epitope Tags

    Type
    Blog Post
    Published
    May 11, 2023, 1:15 p.m.
    ...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...

  23. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    Published
    May 6, 2014, 1:07 p.m.
    ...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...

  24. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    Published
    May 18, 2018, 7:32 p.m.
    ...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...

  25. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...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....

  26. Finding Your Perfect Job After University

    Type
    Blog Post
    Published
    Jan. 12, 2016, 3:30 p.m.
    ...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...

  27. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    Published
    June 23, 2022, 1:15 p.m.
    ...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...

  28. Trading Spaces - Visiting Scientist Edition

    Type
    Blog Post
    Published
    Aug. 22, 2023, 1 p.m.
    ...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...

  29. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ...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...

  30. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

    Type
    Blog Post
    Published
    May 11, 2017, 2:30 p.m.
    ...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...

  31. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...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...

  32. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    Published
    Dec. 15, 2016, 3:57 p.m.
    ...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 ...

  33. Transferable Skills Guide: Problem Solving

    Type
    Blog Post
    Published
    Nov. 30, 2021, 2:15 p.m.
    ...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...

  34. Human Germline Editing Using CRISPR

    Type
    Blog Post
    Published
    Aug. 10, 2017, 2:19 p.m.
    ...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...

  35. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ... 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...

  36. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    Published
    Nov. 14, 2024, 2:15 p.m.
    ...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...

  37. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ...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...

  38. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    Published
    Sept. 17, 2015, 2:30 p.m.
    ...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...

  39. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...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...

  40. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...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. ...

  41. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    Published
    Aug. 21, 2018, 12:31 p.m.
    ...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...

  42. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...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...

  43. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    Published
    Feb. 9, 2023, 2:15 p.m.
    ...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...

  44. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ...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...

  45. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    Published
    Nov. 12, 2020, 2:15 p.m.
    ...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...

  46. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...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...

  47. Career Insights: Technical Support Specialist

    Type
    Blog Post
    Published
    Nov. 9, 2017, 2 p.m.
    ...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...

  48. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    Published
    Oct. 13, 2016, 2:30 p.m.
    ...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...

  49. The Addgene Antibody Data Hub

    Type
    Blog Post
    Published
    Sept. 12, 2024, 1:15 p.m.
    ...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 ...

  50. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    Published
    Oct. 5, 2021, 1:15 p.m.
    ...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...
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