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We narrowed to 948 results for: ache

Showing: 121 - 140 of 948 results

  1. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    ...about the status of each project. This creates accountability on the part of each developer and creates...would be easy to assign each developer to a single area/part of the code and have each developer work only...this can lead to wildly different ideas of how to approach a problem, but it is these very differences that...and the overall soundness of the software design. Each of us brings slightly different cognitive biases...framework is based on an iterative and collaborative approach to development. Scrum teams plan and break up ...mindful of the number of stories we commit to for each sprint, to ensure we will have the capacity to finish...are also able to consider different technical approaches that may come up during discussion.  One example...

  2. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...bearing the "Bunch of Baby Spinach"aptamer, which is a brighter version of Spinach2, an RNA aptamer that fluoresces.... Bunch of Baby Spinach, or BoBS, consists of three tandem copies of the Baby Spinach core embedded in...with Broccoli Vegetable tags, including Spinach, Spinach2, and Broccoli, can benefit your RNA experiments...cleave the target DNA. The inducible split cas9 approach can also be used for activation of specific genes...as TAL effectors or Zinc Finger Nucleases, the Gersbach lab has modified the RNA-guided CRISPR-Cas9 system...light-activated CRISPR-Cas9 effector, Polstein and Gersbach fused the light-inducible protein domains CibN...for many potential applications. Polstein LR & Gersbach CA, Nat Chem Biol 2015 Mar;11(3):198-200. SunTag...

  3. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    ...against this approach. First, this would prevent the study of the proteins independently from each other (see...(6). The EBOV genome contains seven viral genes, each flanked by a transcription start signal sequence...to study minigenomes, these four viral genes are each encoded on separate plasmids and are co-transfected...and the viral mRNAs are translated) by cellular machinery. NP encapsidates the negative sense, single-stranded...all four genes and necessary control elements for each would be relatively large. Cloning the viral genes...this plasmid would be more difficult compared to if each gene were encoded in separate (and thus smaller)...researchers to partially control expression levels of each gene by altering the amount transfected (see Technical...

  4. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    ...complex approaches to scientific questions become popular. There are only so many methods and machines one... a mailing list. This project gave me practice reaching out to other scientists, being clear about what...beyond your organization and/or scientific field. Reaching out to people who have little in common with you...starting a collaboration with a colleague. However, reaching outside your knowledge sphere allows you to connect...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...and started to be fun. I just had to remember to approach the relationship with the perspective of my potential...

  5. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...for a given experimental approach. First, unlike CRISPR, the AAV-based approach is only able to target ...limitations, the AAV approach is best suited for tissue culture based approaches that require modification...working to develop high efficiency gene targeting approaches improved on this finding with the addition of...challenging to develop high efficiency CRISPR-based approaches for the generation of knockins via homologous...sequence-specific knockins - for which the AAV approach is most well suited, since it routinely generates...there are important disadvantages to the AAV-based approach that must be considered when deciding whether ... cell, are more appropriate for a CRISPR-based approach. Second, the AAV preps generally used for gene...

  6. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...directed repairs (HDR). Split AAV vector approaches Split vector approaches increase the size of the gene delivered...following transduction, the partial genomes complement each other to restore the full-expression cassette. AAV... genomes is the basis of several split vector approaches developed in the early 2000’s which allow researchers...splitting the gene into two pieces: part A and part B. Each piece of the gene is then independently packaged...overviews and drawbacks of four split AAV vector approaches. 1. Overlapping  Overview: The overlapping strategy...tested and with the optimal sequences, the group achieved ~80% of muscle cells expressing mini-dystrophin...mini-dystrophin.  3. Hybrid  Overview: The hybrid approach combines the overlapping and trans-splicing methods. The...

  7. Which Fluorescence Microscopy Techniques is Best for Me?

    Type
    Blog Post
    ...These questions will place each sample into one of four categories that are each suited to different types...fluorophores to block out of focus fluorescence from reaching a detector or restricting fluorescence to specific...fluorescent light passes through a pinhole before reaching the detector. The pinhole is specifically placed...placed to only allow light from the focal plane to reach the detector and all light from above or below is...holes rotates through the excitation light path. Each hole corresponds to a different location in the ... the focal volume. This leads to more rapid photobleaching and a reduction in signal intensity. 2-Photon...Photon microscopes restrict excitation (and bleaching) to a single focal point. This is accomplished by ...

  8. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...time, scientists have used almost all available approaches for generating iPSCs. The generation of iPSCs...vectors insert in different genetic locations in each transduced cell, so there’s unavoidable heterogeneity.... Excisable Cre-lox lentiviral vectors: This approach is the same as using a standard lentiviral vector...loxP scar behind at the site of excision, this approach is still not considered safe for therapeutic applications...polycistronic transgene or with four different adenoviruses, each expressing one factor. These vectors don’t integrate...dilution via cell division. A drawback to this approach is it has lower levels of efficiency at generating...transcriptional regulation, so it requires a different approach for regulating transgene expression. Check out...

  9. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...There are several ways to achieve multiplex IF. One of the most common approaches is to use sequential cycles...secondary antibody labeling and imaging, repeating for each target. Alternatively, you can perform simultaneous... 1: Schematic of multiplex immunofluorescence approaches. A) Sequential Multiplex IF, where one target...the sample, either by chemical treatment or photobleaching or by stripping out the antibodies, then start...antibodies are all of different species/isotypes and that each secondary has a different color fluorophore that..., you can still record the signal separately for each target.   Figure 2: Examples of simultaneous...since multiple secondary antibodies will bind to each primary antibody.      Look for primary antibodies...

  10. Antibodies 101: Validation

    Type
    Blog Post
    ...Strategies This approach compares target expression of antibody-independent approaches to your antibody-dependent...Genetic Strategies This approach uses a gene knock-down or knock-out approach to show that an antibody... it’s important to know what you are aiming to achieve. In general, these are three core goals of antibody...antibody-dependent approach. Antibody-independent methods may include analysis of transcriptomic or proteomic...levels of target detected in both methods. These approaches have relatively low sensitivity, which can be... specifically to the expected protein. Common approaches include using RNAi to knock-down expression of...antibody is binding to something unexpected. This approach has high specificity, but you need to be aware...

  11. Bioinformatics at Addgene

    Type
    Blog Post
    ...subsequent process, we linked each step in our pipeline together.  Since each process can use a unique container... expand, we realized the need to modernize our approach to handle larger volumes of data. In this two-...our current need to analyze nearly 300 sequences each week. Our internal software was originally designed...plasmid sequence assembly, and possible solutions for each in the form of one-off scripts or programs, our ...the need to accelerate science using different approaches. We were also encouraged to see that a community...package, all of the pieces of software that depend on each other for the entire package to run. The small, ...complicated setup. We created, tested, and debugged each of our pipeline steps within its own container because...

  12. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

    Type
    Blog Post
    .... Thus, Bernach sought to develop an artificial leaf replica system. “This bottom-up approach will allow... Melissa Chiasson, Elizabeth Unger, and Michal Bernach. Melissa Chiasson: Quantifying the abundance of...based on their eGFP:mCherry ratios. Genomic DNA from each bin are then extracted and sequenced. Chiasson is...vitro against different drugs and dosages.  Michal Bernach: Developing an artificial leaf replica system to...plant-bacteria and bacteria-bacteria interactions  Michal Bernach is a graduate student in Mitja Remus-Emsermann’...Canterbury where he studies how bacteria interact with each other and plant surfaces. His system of choice is...a fully controllable and reproducible set up,” Bernach explains. Developing an artificial leaf replica...

  13. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    ... goal for each analysis will help you locate relevant information in the future. Approach: Outline a...conclusion is “this approach is not ideal because…” Including a conclusion for each analysis or task will...biologists are finding themselves using computational approaches to analyze large data sets (myself included) ...data analyst in our lab took it upon themselves to teach me Python, a programming language favored by biologists... a brief overview of the approach that you will take to help you plan your analysis. There is no need ...scripts and input files that will be used to perform each task. Conclusion: Always, always, write a brief... that lays out which experiments can be found in each folder. This can be a simple .txt document. Record...

  14. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...Mini PCR machines by DIY-Bio, P51 cardboard fluorescence visualizer with 9V battery Approach :This preprint...typical PCR machines, the authors use affordable, field-ready, Bluetooth-enabled PCR machines that can even... required: 37°C heat block and 62°C heat block Approach: Viral RNA is extracted and subject to isothermal... required: 37°C water bath and 42°C water bath Approach: RNA is extracted from a sample and subject to...cleaved. Cleavage results in fluorescence and this approach can be incorporated into a lateral-flow readout...blue light illuminator or UV light illuminator. Approach: Highly similar to the SHERLOCK diagnostic test...temperature. Two Cas12a-crRNA complexes are designed, each binding to one strand of the reverse-transcribed...

  15. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...assembly process designed for a quick, plug-and-play approach to cloning CRISPR vectors (McGee et al., 2024)...design work out of the plasmid creation process. Each module of Fragmid represents a class of components...enzyme or Pol 2 promoter. The options available in each class, or module, are the components. Using the ...users can simply select the components they want for each module, generate the vector file, and quickly clone... 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 ...outside of its recognition sequence. This means that each component can be designed with an overhang specific...

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

    Type
    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... Delivery of AAV into the brain can usually be achieved by a directed brain injection, but some serotypes.../loxP or Flp/FRT, with AAV vectors are common approaches to control transgene expression by restricting...types. Specific transgene expression can be also achieved in nontransgenic animals by using cis regulatory...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...

  17. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ... executive director of Central Indiana Science Outreach and a Postdoc at Indiana University School of ...Biology, I realized I wanted a career in science outreach: engaging the public to promote an awareness and...founding a nonprofit called Central Indiana Science Outreach, or CINSO. We organize fun science events for ...do fun science activities with the public, so I reached out to people who were educating the public and...and learned a bit about recruiting scientists to outreach.  Try new things to further your career Take advantage...new people. There were visiting seminar speakers each week, my department and graduate office arranged...Especially as someone interested in doing science outreach, not only can I easily share science with the ...

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

    Type
    Blog Post
    ...resulting in processing by the endogenous RNAi machinery. Alternatively, endogenous miRNA precursors can... either technology was equivalently precise, but each method recovered different genes known to be required... two. Consequently, I will focus on different approaches and considerations present at the first stage...initial state of the dsRNA upon activating the RNAi machinery (long dsRNA, typically hairpin RNA, or artificial...long-term RNAi phenotype. Long hairpin RNA The easiest approach to induce RNAi involves the expression of long...different constructs are available to the community. Each construct has unique features, such as inducible...RNA The greatest limitations of the long hpRNA approach are thought to be off-target effects and variable...

  19. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ... passes through the machine’s lasers and gives off a signal read by the machine’s sensors. (Usually this...could be debris or unbound antibodies.) Each event read by the machine is plotted as a dot on an X,Y axis ...plotted on an invisible Cartesian plane (right.) Each event is plotted by the amount of signal read from...variations in fluorescent signals, and how the machine reads/interprets the signals, the axis is instead...populations. This shape is called a ‘gate’; it tells the machine that everything inside the gate is one population...doublets and single cell populations are very close to each other.        Figure 4: Gating strategy ... numbers representing the percentage of cells in each gate. Used under Creative Commons license.  ...

  20. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    ...single model. The first approach can lead to new directions, while the second approach could terminate with...the problem and goal, the approach, anticipated outcome, and payoff for each aim. Learn to communicate...preparing for both types. Your research vision is your approach to resolving a significant knowledge gap in your... you need to be able to describe experimental approaches in terms of the question, the methods, the tools...question, the anticipated outcomes, the alternative approaches, and what will you do if your model is correct..., a clear easy-to-understand question/goal, an approach, anticipated outcomes, and preferably a description...successful. Note that aims do not need to be grant aims. Each aim could be the basis of a single grant. Either...
Showing: 121 - 140 of 948 results