...section.   Additional Resources on the Addgene Blog Apps for Lab Life Developing Lab Management Software ...

...titles and abstracts are available online or in an app prior to the session, so bookmark which posters are...background without an introduction. If someone approaches your poster, introduce yourself and ask about... a related topic, and let them know while you appreciate the thought-provoking question, you are unable... enjoy hearing the highlights so that you can appreciate the impact of the project as a whole. In addition...

... scientists––CrispyCrunch! CrispyCrunch is a web app that helps scientists design and analyze batches ...score is less than 0.03, it stops mutating. (This happens often because the HDR inserted sequence splits ...Psychology at McMaster University in Canada. He's happy to be contributing again to science.   Acknowledgements...

...letters to be very important. On the other hand, our app/dev team rarely, if ever, sees cover letters submitted...usually yes! A good cover letter won’t hurt your application, but it can certainly help you stand out.  In...consider cover letters an important part of the application. Even within an organization, there can be variance...good cover letter!) As with any part of the job application process, it's a good idea to talk to people in...do the job as described. Let’s imagine you’re applying to a job that requires ddPCR skills. Your resume... to proactively address questions about your application. These could be logistical, like briefly explaining...  Finally, it’s a good idea to state why you’re applying to the specific company or lab. Take some time...

...Learn about more tools for reproducibility 5 great apps for lab life Developing Lab Management Software ...sciNote. Today, every lab has its own habits and approaches to record keeping. Top priorities in most cases...Project: Cancer Biology have shown similarly disappointing rates of reproducibility. These alarming results...familiar paper-based lab notebook is still a standard approach to record-keeping in many labs. In a way, it will...digital data generated in labs. While many labs approach the situation differently, some adopting simple...might be available as well. In most cases this applies to universities. How are you currently exchanging... Besides this being of crucial importance when applying for a patent for example, it also comes in handy...

...mutations in genes associated with dementia, such as APP and PSEN1, in most cases the intended HDR events ...with blocking mutations, we still weren’t entirely happy with our editing outcomes and in many cases still...

...tools as some kind of a meta/Frankenstein app? Or perhaps there is a clear winner, is there something that...the one hand we have software tools that apply some approximate or sensible rules to determine the value... more efficient.  These insights are hard to appreciate without computational support. Indeed, rational... solutions that facilitate most experimental applications in CRISPR engineering. When compared to that...easy to fault the lack of features in a single application. The question is then, should we use all tools...2013) used a combination of both empirical and approximate methods. The details of their scoring function...and it means that there is no one-size-fits-all approach. You will have to select a tool based on your ...

...single assay. Overview of Antibody Applications Antibody-based applications can be generally classed into ...these applications, visit the Antibody section of the Addgene Protocols page. Antibody Applications - Quantification...of signaling molecules. Select one appropriate for your application and experimental design. Will you ...transcriptomics or proteomics. This approach is often appropriate for quantitative methods. Capture MS...protein of interest, with no overlapping epitopes. It is not appropriate for validation of an antibody...production and storage techniques, and explain common applications. Science... Handling Visualization Signal Amplification Application Overview Quantification Methods Capture Methods...

...26628643 2015 A Scalable Genome-Editing-Based Approach for Mapping Multiprotein Complexes in Human Cells. 2015...engineering . Prior to CRISPR, genome engineering approaches like zinc finger nucleases (ZFNs) or transcription-activator-like...rapidly become the most popular genome engineering approach. Engineered CRISPR systems contain two components...understanding of CRISPR biology, introduce the various applications of CRISPR, and help you get started using CRISPR...experiment, you might consider using the dual nickase approach to create a double nick-induced DSB. The nickase...have been engineered using cell-based selection approaches. Yeast-based screening systems resulted in evolved...requiring longer homology arms. Depending on the application, the repair template may be a single-stranded...

...library. Apply pooled library to cells. DIVERGE: Positive and negative screens POSITIVE screen: Apply library...control sample (no selection) In parallel, apply library Apply selection; most cells will survive Perform..., semi-random sequences that can be used for applications like lineage tracing or parsing the effects ...library Apply selection Most cells die or do not pass selection (in the case of a reporter) Sort “winning...genes involved in the selection NEGATIVE screen: Apply library Perform Next Generation Sequencing (NGS)...and control cells Generate a list of gRNAs that disappear with addition of the selection mechanism Positive...

...move around pieces of DNA. Oftentimes several approaches will work for any specific cloning project; however...likely that for any given project there is an ideal approach. This may be due to speed, cost, availability ... of time required for cloning. To utilize this approach, the fragment of DNA that you would like to clone...fragment (PCR product or synthesized oligo) with appropriate overhangs can be efficiently ligated into any... enzyme cloning. Scientists have utilized this approach to create compatible custom overhangs that can...fragments of dsDNA that have 30 or more bases of overlapping homology. One major advantage is that much larger...

...assume that their advisor alone applies for these competitive grant applications; however, in some cases, your...the grant application process. When my PhD advisor first asked me to write a grant application due the ...mind that universities require application submission in-house approximately 2 weeks before the RFA due date...the next round of applications. Step 5: Keep writing! Each written grant application builds upon the ... project grant application" by the NIH Still want more information on the application process? What types...writing, volunteered to take the lead on various applications throughout my graduate career, and helped receive...opportunities to convince others why a project is the best approach to a given problem, Empowers the writer, And...

...intended application - Remember, just because your antibody performs as expected in one application does ...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...protein. This approach is ideal for validating antibodies for immunocapture applications and is amenable...pause and make sure that your antibody has been appropriately validated so that you can have confidence in...in your results. But what is appropriately validated? This post will go over some of the general considerations...strategies for antibody validation in different applications. Here, I want to highlight the general suggestions...

...therapeutic applications. Clinical trials using AAV for various gene delivery applications are currently... preferred over standard viruses for in vivo applications. Scientists have also experimented with hybrid...transgenes, enabling the development of further AAV applications. Return to Top Additional Resources AAV Plasmids...AAV packaging capacity with trans-splicing or overlapping vectors: a quantitative comparison. Duan D, Yue...

...allow you to approach that problem in a novel way. Finally, engineering new applications to solve problems...good quality application. Even following this advice, only about 50% of my grant applications have been ...do need to be able to research and write your application, then give it to someone else to read and / or...it). It’s easy for reviewers to spot when an application is rushed (whether the idea isn’t properly researched...researched / developed or the application is full of typos) and these are unlikely to get past the initial...someone else in the department) to read over your application and give you feedback. They’ve most likely been...alterations before submitting - handing them your application 3 days before the deadline isn’t going to be ...

...of four split AAV vector approaches. 1. Overlapping  Overview: The overlapping strategy has a region of...directed repairs (HDR). Split AAV vector approaches Split vector approaches increase the size of the gene delivered...  3. Hybrid  Overview: The hybrid approach combines the overlapping and trans-splicing methods. The AAV...has a slight advantage over the overlapping and trans-splicing approaches since there are two ways for a... genomes is the basis of several split vector approaches developed in the early 2000’s which allow researchers...is not yet known. Drawbacks:  Generally an overlapping split vector has equal or higher transgene expression... on the rate of transgene expression with an overlapping AAV since both of these variables influence whether...

...is this application - the creation of sequence-specific knockins - for which the AAV approach is most ...for a given experimental approach. First, unlike CRISPR, the AAV-based approach is only able to target ...simultaneously in the same cell, are more appropriate for a CRISPR-based approach. Second, the AAV preps generally...limitations, the AAV approach is best suited for tissue culture based approaches that require modification...researchers could apply this technique to efficiently deliver large repair templates for applications that require...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...

...neurons) that can be turned on or off by the application of a small molecule ligand. The ideal chemogenetic...translational studies because it is FDA and EMA approved. One pitfall of a suite of muscarinic-based DREADDs... develop a PSAM that responded to a clinically approved drug, the PSAM domain was engineered to respond...Azua IR, Li JH, et al (2009). A chemical-genetic approach to study G protein regulation of β cell function...chemogenetics for research and potential clinical applications. Science: 364(6436):eaav5282. PMID: 30872534...

.... This is the direct approach to immunoimaging assays. But if the direct approach isn’t giving you enough...   Figure 1: The direct approach (left) versus the indirect approach (right). Primary antibodies ...varies across applications. Western blots, for instance, almost always use the indirect approach, while flow...immunocytochemistry may all use either the direct approach or indirect approach, depending on the requirements of the...interest, it may be time to look at the indirect approach - which means it’s time to talk about secondary...binds to the primary antibody. In the indirect approach, the signaling molecule is conjugated to the secondary...antibodies can bind to a single primary antibody, this approach can greatly increase the sensitivity of an assay...

...vector, some possible approaches to using this site include subcloning or appending compatible restriction...packaging and envelope plasmids are generalized and appropriate for varied cell types and systems. When planning...generation systems) Safety Concerns Lentiviral-based applications Read our detailed protocol for using the popular...begins, by definition, at the beginning of R, is capped, and proceeds through U5 and the rest of the provirus...