We narrowed to 953 results for: APP
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TypeBlog Post... can happen at the national level within the executive branch or in Congress, or can also happen at the... there will be posts about virtual meet ups and happy hours that you can attend to help build relationships...arriving here in DC, is that so much policy actually happens outside of the government itself. You can work ...The “little p” policy is non-governmental work happening at large organizations who have to develop policies...who decides how they are going pursue regulatory approval for a new product or a well-endowed philanthropy...Program to learn more about where science policy happens. When do you do science policy? Many science ...Project for some example proposals and consider applying for their accelerator for some hands-on guidance...
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Negotiating Work and Life: How to Find the Joy
TypeBlog Post...sense and appreciate this – and they will be happy too. On the flip side, if you are not happy at work,... If you are happy at work, you will find balance. If you enjoy working and come home happy in the evening...actions we can take to help ourselves can be an appropriate first step. Check out Joanne's Reddit AMA Managing...turns going; that way you can have someone there clapping more often than not. You want your family and ...tools and tactics to plan your work and make this happen. Be realistic and: Your co-workers will learn ... about their safety when they leave home which happens very, very fast! I will never forget the time my...figure out a way to keep being ourselves and to be happy. Building your plan to live a good life may be the... -
Overcoming the AAV Size Limitation for CRISPR Delivery
TypeBlog Post... are approximately 4.1 kb in length, allowing them to be packaged in AAV. Split AAV approaches While...germline genome editing, but in vivo gene editing approaches have been limited by problems with Cas9 delivery...packaging capacity of AAV (~4.7 kb). While this approach has been proven feasible, it leaves little room...potent Cas9 protein, the Zhang lab took a different approach. They analyzed over 600 Cas9 orthologs and found...be divided into two groups: longer orthologs approximately 1350 amino acids in size, which includes SpCas9...SpCas9, and shortern orthologs approximately 1000 amino acids in size. From the pool of shorter orthologs...effects of SpCas9 and SaCas9, Zhang’s group used an approach called BLESS (direct in situ breaks labeling, ... -
Hot Plasmids - November 2023
TypeBlog Post...platform offers a promising approach for experimental therapies and medical applications. Find DIRECTED plasmids...Figure 1: Choosing the best PE6 variant for your application. Image reused form Doman and Pandey et al., 2023...REcipient Cells Through Envelope Design). This modular approach combines an interchangeable set of fusogenic and... CreVLPs, and eVLPs for in vivo and in vitro applications. Figure 2: Modular features of DIRECTED...interactions or map a local proteome, but some applications have been challenging or inaccessible. The Ting... Qin, W., Cheah, J.S., et al. (2023). Dynamic mapping of proteome trafficking within and between living... from different promoters, specific time points appear as fluorescent bands in the fiber, resembling a... -
SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas
TypeBlog Post...fluorescence can be visualised. Another approach was to apply the same concept to lateral flow strips ... Suresh from Iowa State University. Since its appearance, SARS-CoV-2 has spread to almost every part of... 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...microliter Time: Not mentioned in the paper, likely approximately 1-2 hours Equipment needed: Mini PCR machines... -
Antibodies 101: Introduction to Immunofluorescence
TypeBlog Post...is an application in which antibodies that have been joined to fluorescent molecules are applied to a ...immunofluorescence There are a couple different approaches to be familiar with when talking about IF, direct...BioRender.com Indirect IF is a much more common approach than direct IF, so we will focus on this technique... Primary Antibody This is the first antibody applied to your sample, which will bind to your target ...Unsurprisingly, this is the second antibody you apply, which binds to the first antibody and is conjugated...distribute antibodies validate them in a variety of applications, including IF (or IHC or ICC), and will share... time to research antibodies, protocols, and appropriate controls and you will be well on your way to ... -
Antibodies 101: Normalization and Loading Controls for Western Blots
TypeBlog Post...sample of your protein of interest (POI). This approach is reliable, but it’s overkill in many cases. ...than “condition 2.” By eye, the loading control appears about the same between the two conditions. But ...detected with an antibodies, just like the POI. This approach is so common that we’ve used it for our cartoon...cells. This means that at sample concentrations appropriate to detect a less-abundant POI, the loading control... control aren’t too far apart, they may have overlapping linear detection ranges, allowing you to accurately..., 2008; Gilda & Gomes, 2013). Depending on the approach you choose, your loading control measurement will...on the same gel or membrane (depending on your approach) as your final blot. Quantifying the entire lane... -
Plasmids 101: Methylation and Restriction Enzymes
TypeBlog Post...Blocked by overlapping methylation Not affected BsaI Not affected Blocked by overlapping methylation...Blocked by overlapping methylation HpaI Not affected Not affected Blocked by overlapping methylation...Blocked by overlapping methylation Not affected Not affected MscI Not affected Blocked by overlapping methylation...Not affected Blocked by overlapping methylation XbaI Blocked by overlapping methylation Not affected...methylation Not affected ClaI Blocked by overlapping methylation Not affected Not affected DraI Not affected... -
Is this the right place for me? 8 tactics for choosing a lab
TypeBlog Post...think “but that won’t happen to me, I have a backbone”. The sad truth is, if it happened before, it almost...choosing a lab, but almost all of these guidelines can apply to any workplace or job. It amazes me when the most...scheduled or only ad hoc? Does the mentor keep appointments with group members or are they frequently cancelled...conflicts in the lab? Do they motivate with the appropriate balance of encouragement and criticism? Do they...was told that the PI removed a woman from the appropriate first authorship position on a publication of...and profoundly regretted it when the same thing happened to her a year later. I hear too many science trainees...almost certainly will also happen to you. As you speak to lab members and alumni, don’t ever disregard warning... -
Transferable Skills Guide: Public Speaking
TypeBlog Post...t just happen to you or me (though, let's be clear, they have all happened to me.) They happen to everyone...is, when it happens to other people, it’s normal and quickly forgotten, and when it happens to you, it’...communicator, I’ve done a lot of public speaking, and I'm happy to share what I've learned along the way. Own your...comes to public speaking, Murphy’s law definitely applies: what can go wrong, will go wrong. The trash removal...interest incorrectly on slides 8-16. Your notes disappear. The wrong version of your slide deck gets uploaded...completely normal to be anxious, nervous, and/or apprehensive. I still get nervous quite regularly, and I'... -
27 Hot Plasmids from 2016
TypeBlog Post...AID an appealing tool for studying essential proteins. Until recently, it was challenging to apply the AID-tagging...brief articles highlight the main features and applications of a partiular plasmid or set of plasmids. We...December 2016 (Q3) MARCH 2016Enhancer activity mapping with STARR-seq Although many technologies have...classical TANGO assay, Kroeze et al. developed a new approach termed Parallel Receptor-ome Expression and Screening... (PRESTO-TANGO) to screen the NCC-1 library of approved drugs against the entire kit; a parallel analysis...multiple expression systems before they achieve the appropriate expression level for their gene of interest. ...GMAP-compatible vectors for a wide variety of biological applications. First, GMAP-compatible backbones for lentivirus... -
CRISPR in the Clinic
TypeBlog Post... drug approval boards are being finalized, and it is very likely the treatments will be approved for use...disease. Meanwhile, Hans Clevers and colleagues applied CRISPR to disease correction in adult stem cells...CRISPR-based editing therapies that are currently approved or have made significant progress in clinical ...Therapies Cancer and CAR-T therapy The FDA has already approved CAR-T immunotherapy for cancer treatment by traditional...These therapies are delivered with an autologous approach, editing patient cells ex vivo and then returning...but especially concerning when using a systemic approach. Regarding effectiveness, if the editing frequency...meaningful way. Delivery, especially for systemic approaches, has its own unique set of obstacles. Thus far... -
FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution
TypeBlog Post...which subpopulation of DA neurons could activate appetitive rather than aversive stimuli. In a recent paper...rAAV FLEx-based vectors to answer this question, mapping and characterizing the activity of the DA neurons...pursue a teaching career, and she now is a very happy scientific communication manager at genOway. ... cell types for imaging and long-range circuit mapping." Journal of Neuroscience 28.28 (2008): 7025-7030...Philippe Coulon, and Ilka Diester. "Optogenetic approaches to study the mammalian brain." Current opinion...Yizhar, and Karl Deisseroth. "The development and application of optogenetics." Annual review of neuroscience...Mei, Yuan, and Feng Zhang. "Molecular tools and approaches for optogenetics." Biological psychiatry 71.12... -
Troubleshooting and Optimizing a Western Blot
TypeBlog Post...Alternatively, you can run a reagent gradient. In this approach, you’ll run the same protein concentration in ...concentration of your reagent. Be aware that this approach requires excellent attention to detail, since ...down to voltage and time. Make sure to load an appropriate protein ladder on your gel and keep an eye on...too high. If you’re seeing a smiley face at an appropriate voltage (usually 10–15 V/cm of gel), your gel...box. If overheating remains an issue, which can happen with large proteins, try lowering the voltage and... buffer — so it’s better to start over if that happens. Finally, if you’re seeing a weird issue and can...your reversible staining method of choice is appropriately sensitive. If using a PVDF membrane, you can... -
When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells
TypeBlog Post...Many thanks to our guest blogger Erik L. Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite...problem for fusion proteins. One of the major applications of FPs is to visualize the localization, dynamics... dimerize can form dynamic contacts that bring apposing membranes together and can wrap the membranes ...the Endoplasmic Reticulum and live cell imaging approaches. Erik is also an avid long distance runner, gardner... gardner, and cook. References 1. Snapp, E.L., et al., Formation of stacked ER cisternae by low affinity...An experimental study of GFP-based FRET, with application to intrinsically unstructured proteins. Protein... -
CRISPR 101: Mammalian Expression Systems and Delivery Methods
TypeBlog Post... the appropriate CRISPR tools for your application: Wildtype Cas9 or Cas9 nickase are appropriate for ...well as suitable applications. Expression System Components of System Application Mammalian expression...CRISPR mammalian expression systems, the typical applications for each, and potential delivery methods. ...mutations near the gRNA target site. Choose an appropriate expression system and delivery method: Do you...electrophoresis and/or next-generation sequencing more appropriate? If you already have an end product in mind...or more sgRNAs and Cas9 are required for any application. If you want to include a homology directed repair... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog Post...amenable to transfection or viral transduction Appropriate promoters must be chosen for both Cas9 and gRNA... Cas9-gRNA ribonucleoproteins? One alternative approach, which avoids many of these complications, is ...used for most of the current genome engineering applications of CRISPR including: generating single or multi-gene...pathways. This makes Cas9 RNPs useful for CRISPR applications where limited expression of Cas9 is required...vendor, this can often be expensive. An alternative approach is to express and purify His-tagged Cas9 from ...this technique in cell culture, it has also been applied to genome editing of mouse zygotes, through a process...inner ear (Zuris et al. 2015). A more targeted approach using RNPs has recently been described in vitro... -
MXS Chaining
TypeBlog Post...engineer various constructs with applications in flow cytometry approaches in mammalian cell culture systems...develop diverse fluorescent reporters is a useful application of high-throughput cloning. MXS Chaining is one...plasmids for fluorescence imaging or flow cytometry applications in mammalian cells. The modules used in this...transgene expression, or a variety of other clever applications. In one example, modular constructs generated...methods are suited for particular downstream applications. The tradeoffs between various cloning methods...from them. These enzymes might not, however, be appropriate for cloning genes from organisms with high CpG...quickly, and simply. We hope that you can similarly apply MXS chaining to your experimental needs. Let us ... -
CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics
TypeBlog Post...structural proteins that make it possible, see the appendix DNA is a 3D fractal). Enhancer biology is a complex...acetylation) - tri_methylation In the next section we’ll apply the above formula to a real-world experiment. We...: the enhancers! This leads us to a three-step approach for designing an experiment to identify the critical...therapeutics. We’ll have more to say about clinical applications of CRISPR in the next section. In order to denote...mechanism for CRISPR. Therefore if a gRNA drops or disappears over time, we infer that the enhancer it targets...own CRISPR screens for enhancers, check out the Appendix. Conclusion: Shedding light on dark matter Precise...is the foundation of precision medicine. We can apply this knowledge to create targeted disease therapies... -
A Career in Grants: How To Become a Grant Professional
TypeBlog Post...the candidate application process, the candidate’s ability to write a grant application through a written...funding opportunity coincided, and they stepped up to apply for a grant which lead to their new career path.... much more than writing. In fact, many grant applications and grant makers have moved away from long written...Now they require grant seekers to use online applications with significant character limits for the written...written portion of the grant application. They have increased their demand for stronger budgets with clear...duties can range from coordinating the grant application and submitting it, to evaluating the capacity...grant readiness, funding opportunity research, application writing and development, project implementation...