...sequence? Do these two proteins interact? What happens when I overexpress this enzyme at a particular ...cellular activity in a way that’s interesting from an applied perspective, sometimes it’s best to delete genes...bacteria of interest are transformed with the appropriate construct, grown at the permissive temperature...localization of your gene of interest once its appropriate cDNA has been fused to the MS2 binding sites ...

... generation CRISPR-Cas platform for multiple applications....by the PUF code, it is possible to construct approximately 60,000 orthogonal modules that can function ...combinations of modules as is, allowing plug-and-play application of Casilio in functional genomics studies. Overview... sgRNA-PBS plasmids A single-guide RNA (sgRNA) appended with multiple copies of Pumilio binding sites ...

...sure I missed some of your favorite apps. If you know of a software app that I missed, let me know in the... The software can be used with the ZappyLabs Bench Tools mobile app (available for iOS and Android). A...system does not appear to have a way to share your protocols with other groups. What apps do you use to ...publishers, and small start-ups are turning to software apps to solve our need for quicker, more effective updating...more to fit every lab's needs. Plus there are the apps - many to help you access your protocols at the ...comments below and leave your thoughts on how the app works for your lab’s needs. At Addgene, we’ll continue... some of our basic protocols to the protocols.io app so that all of our researchers can easily take those...

...have been developed for different scientific applications. These PCR methods all use the same basic PCR...quantified by qPCR. qPCR is used in a variety of applications including gene expression profiling, studying...that fractions the PCR reaction sample into approximately 20,000 droplets. Each droplet contains the material...

...their use in other clinical applications. In the last decade, viral applications in cancer treatment and ...s unpack the myriad number of emerging viral applications and give you some recommendations for putting...putting viruses to work. Viral applications Viruses work by infecting a host cell (the target of viral infection...expressed. It’s important to note that for all the applications we discuss, there is more than one type of virus...retrovirus, AAV, etc.). When you’ve decided on an application, our Viruses as Biological Tools blog can help...viruses and pick one for your experiment. Research applications Gene expression control Researchers are always...becoming increasingly popular tools with many applications. These libraries include sgRNAs targeting the...

...options, you’re likely to find an application that matches your needs. Happy quantifying!  References and...blog post, we're going to focus on PCR-based applications (qPCR and ddPCR.) qPCR Quantitative/real-time...range of RQ values as noted above.   In both applications, results can be affected by DNA quality and ...as positive or negative for each reaction, math happens (this time, Poisson statistics), and the readout... triplicates. ddPCR is an extremely accurate application, even in areas of the genome that are difficult...experiments, a number of factors will determine which approach is right for your experiment. You’ll have to consider...QuantificationPolymerase Chain Reaction Overview and Applications Resources on Addgene.org Protocol: Polymerase...

...particular application can be found here.  Why CRISPRa? Although cDNA overexpression approaches may still... viral vectors, and high-throughput screening approaches. Here, we’ll highlight these recent advancements...advancements and offer commentary on their application to neuroscience research. What is CRISPR-based activation... a larger packaging capacity than AAV. Such an approach was described recently by the Day Lab, who developed...accessible to neuroscientists, not all neuroscience applications require such optimized tools. This is elegantly... developed a subgenome-scale CRISPRa screening approach in embryonic stem (ES) cells to determine which...study neuronal differentiation, it could also be applied to the differentiation of other CNS cell types,...

...and applications.    Plant applications We outlined many synthetic biology MoClo applications, but...for synthetic biology applications in bacteria or cell-free systems. The approach is particularly useful...- and multi-gene constructs for a variety of applications. MoClo tools can be used in bacterial and cell-free...and the kits Addgene offers to kickstart these applications, no matter what your model organism is. MoClo...this blog, we will highlight some common MoClo applications and kits. Keep in mind that this technology ... it, MoClo might help you achieve it! MoClo applications and specialized tools Synthetic biology in bacteria...effectiveness. Perhaps the most intuitive CRISPR MoClo application is to assemble multiple unique sgRNAs into a ...

...function (Wood, et al). In the microbial world, approximately 50% of predicted genes have unidentified roles...endogenous gene. Each trap uses a slightly different approach (described in detail below), but fundamentally...redundant genes – multiple genes which function in overlapping roles can mask the depletion of one another. ...trap systems lack a promoter and thus require the trapping of an endogenous promoter. The system is composed...enhancer traps lack enhancer elements and require the trapping of endogenous enhancers to stimulate the expression...many more! Check out Addgene’s catalog to find trapping tools for your next experiment! References and...

...females at an incidence of approximately 1:10,000. It is characterized by apparently normal development through...In females, random X inactivation results in approximately half of cells expressing wild-type MECP2 and...Tools CRISPR technology is a promising therapeutic approach for precise editing of mutations in the MECP2 ...

...protein for your application. Selecting the best fluorescent protein for a specific application can seem like...under conditions that best mimic the intended application. The three key properties for live cell imaging...theoretical brightness. The theoretical brightness often appears in tables with fluorescent protein properties and...experimental set up. The practical brightness takes all application specific parameters into account, including the... picture of what can be expected in a 'real' application than the theoretical brightness. Consequently...brightness in a system closely mimicking the future application. In figure 2 we provide an example of the comparison...

...cas9 gene and single guide RNAs (sgRNAs) with appropriate flanking homology into a germline cell can create...RNA-guided gene drive. In principle, this event could happen even if there isn't any homology available. Consequently...local biosafety officer should be familiar with appropriate measures. Barriers should be a component of all...eventual release should consider building an appropriate immunizing reversal drive at the same time to...species in question), why not use them whenever applicable?Against this, consider the cost of an accidental...An accidental release could delay real-world applications against scourges such as malaria and dengue .... Disclaimer: Recommendations concerning the appropriate degree of confinement are those of the author...

...The Framework for In vivo T cell Screens (FITS) approach (Milling and Markson et al., 2024), developed ...compatible with single gene KO and pooled screening approaches; (4) encodes a Thy1.1 selectable marker to facilitate... et al., 2024.   Proof of concept The FITS approach was used to optimize conditions for a screen that...

...explore alternate approaches. Being a “lazy” geneticist, I developed a co-selection approach relying on repair...with unprecedented ease. These new C. elegans approaches rapidly enrich for editing events without the...phenotypes (unc-119) (Dickinson et al., 2013). These approaches allowed effective recovery of knock-ins, but ...Including Cre-mediated marker excision, it takes approximately 3-4 weeks to obtain a homozygous, outcrossed...several manuscripts published in Genetics detailed approaches in which selection for an editing event that ...and knock-ins at other genomic loci. The first approach – co-CRISPR from Craig Mello’s lab – used inactivation...animals with particular visible phenotypes. These approaches may become even more powerful following a report...

...advantages and disadvantages depending on the application. Firefly luciferase is considerably brighter ...luciferase-based plasmid systems for many different applications. Browse the Collection Highlights section to...respectively, for increased usefulness in in vivo applications. LuxSit-i : An artificial luciferase desined...desined de novo using deep-learning approaches. Split-Luciferase and Protein-Protein Interaction Assays Cluc-FKBP...

...transduction (detected by fluorescent expression when applicable) PCR-based confirmation of the viral genome Full...submitted an AAV Made on Request inquiry, now what happens? Once a Made on Request inquiry is submitted the...feasible. Please note that we cannot guarantee legal approval to prepare a plasmid as viral particles. A member...

...antibodies are produced in-house and undergo application-specific validation and quality control by Addgene... Recombinant Antibody Reactivity Recommended Applications PI Don’t See What You’re Looking For? Please...email to [email protected] . After we review and approve the MTA documents, your order will be processed...

... the app. Technical difficulties? Attending a conference virtually means getting to know the app or virtual...conferences with multiple sessions happening at once, virtual conferences may have apps or interactive calendars...was able to get in touch with the author. “In the app, you can write a message to the author of the poster...there may be a “place” for networking within the app. For example, the NeuroSymposium had virtual tables... puts butterflies in your stomach, the virtual approach might be less stressful. For both conferences,...Having an on-site conference canceled may be disappointing, but there are many creative ways to adapt and...

...useful to help improve this technology or its application? What are some current obstacles that researchers...antibodies and recommended dilutions, etc.), novel applications, or any other useful information. Experimental...take care of that. Note that you will have final approval before anything is released publicly, but please...