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... allows components to be swapped out, potentially broadening its applications. Find ATLAS plasmids here...Addgene’s own AAV6 and AAV11 packaging plasmids Mapping anterograde synaptic connections with ATLAS Bright...between mGold2s and mGold2t depending on the application. Figure 1: Super-resolution imaging...proof-of-concept, they believe that the strategy will be applicable with other TnpBs and in other plant species, ...available for tracking cell lineage, different applications require careful selection or design to ensure...Alejo Rodriguez-Fraticelli recently developed an approach called Simultaneous Tracking of Recombinase Activation...BY license. Libraries are available with T-Sapphire, EGFP, or mScarlet reporters, so you can choose...

... to be placed immediately upon request approval. Once approved, the request will remain in your account... in this blog post.     When will the expansion happen?  You will be able to request Packaged on Request...request will be reviewed by Addgene for legal approvals and technical compatibility with our production...process will take 1–2 days, and we anticipate approving the vast majority of requests.  If the vector ... request process.  Ordering If your request is approved, you will receive an email notification that your...is ready to order. An “Add to Cart” button will appear with the request in your Addgene account.  Orders...viral preps will be available at launch.  What happens after I order?  Once an order is placed, the Addgene...

...distinction of neighboring cells in imaging applications and the mapping of neuronal projections to their associated...while minimizing confounding secondary effects? An approach described as “knocksideways” (a British idiom ...domains, such as FKBP and FRB. The cell does not appear to mind extraneous proteins bound to its outer ...protein from expression to degradation. Chemical approaches may require expensive peptides or small molecules...and Technology Austria, decided to take a novel approach to control for these effects, drawing on the rapidly...optogenetics. Using a rational protein engineering approach, they designed Opto-RTKs, which activate signaling... on exposure to low-intensity blue light. This approach relies on the incorporation of light-oxygen-voltage...

... 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...

...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...

...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...

... 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, ...

...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...

...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 ...

...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...

...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...

...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...

...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'...

...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...

... 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...

...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...

...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...

...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...

... 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...

...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 ...