...properties make nanobodies easier to transform and express in bacteria, which, in turn, makes them more...summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you...

... A good rule of thumb is to find the pieces that make you think, “Oh, that’s interesting! How did you ...in with something that sparks their interest and makes them want to know more. This could be by outright...needs to capture your audience’s attention enough to make them decide to read (or listen or watch) further...

...resolution functional tracking of T lymphocytes, too, making them a versatile tool for application in both neuroscience...element-binding protein (CREB) in living cells. “Now we are making this really good,” Griesbeck said. “We know CREB... but not the other). “It’s quite a robust way to make a FRET sensor with a large dynamic range,” he says...

...researchers to utilize and develop these tools by making their own fluorescent protein technologies widely...off with light.  Xin hypothesized that she could make enzymes controllable by tethering them to Dronpa...communication skills throughout her training and wants to make her work understandable to people both inside and...

...express YFG in a new tissue type (more details on making a fly line in the next blog post). For example,...get the right cell type expression as opposed to making a new fly with hedgehog under a new promoter each... are already available, sometimes you’ll have to make your own. In my next post, I’ll explain the relatively...

...Professor at University of Barcelona. The challenge: Making courses on genetic manipulation more hands on While...University of Barcelona wanted to develop strategies to make our classes more practical and hands-on. Students...interactive setting. The problem is…how can you make a “hands-on” lesson on genetic manipulation of human...

...greater familiarity with scientific methodology makes it easier to spot even minor weakness or limitations...Zealand’s multiple Covid-19 lockdowns, I held mask-making workshops in my community. I brought along printed...run workshops where I showed people how they can make reusable cloth menstrual pads while we talked about...

... of making science easier and more transparent. That’s largely my job [at the Broad] too… to make science...help guide and promote Addgene in its mission to make research easier. Addgene is excited to announce ...

... Therefore, it is not always mission critical to make sure the plasmid replicates in the cell, as many... (i.e. the plasmid contains a cassette that will make cells resistant to a toxin), whereas negative selection... (i.e. the plasmid contains a cassette that will make cells sensitive to a toxin). In the table below ...

...  The antibody-producing B cells that are used to make hybridomas come from animals that were immunized...ligand during your experiment, then you’d want to make sure that you choose an antibody that recognizes...disadvantages of monoclonal antibodies will help you make the best decision for your experimental needs.   ...

.... Before you read out your plots, you’ll want to make sure you’ve also adjusted voltage and compensated...tips on adjusting voltage: During data acquisition, make sure that the signal for the positive population... dot plot in Figure 2. Quantification using MFI makes it easier to identify and study these subtle shifts...

...their precision editing capabilities. Base editors make specific DNA base changes and consist of a catalytically...They may be slow, can only target certain sites, or make only a subset of base substitutions.  The need ... to the base editing toolbox, scientists can now make these edits much more efficiently and in more contexts...

...promise there is the ability share the information, making it available to everyone, that creates more breakthrough...looking for those “what could go wrong scenarios” and making sure we have adequate process and controls in place... in a particular area or profession, you need to make sure you have learned and mastered the fundamentals...

...summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you...however can interfere with other genetic pathways making it difficult to link a phenotype to the gene being...The increased efficiency of TurboID and miniTurbo makes PL a viable method for proteomics in model organisms...

... continues to make improvements to our website and inventory management systems to make way in our systems...plasmids in the repository. The Outreach Team has been making sure scientists know about the new initiative and...

...head-to-head comparison of fluorescent proteins and how to make a well-informed decision on the best fluorescent...properties of fluorescent proteins on websites. To make the best choice, the fluorescent protein properties...than the theoretical brightness. Consequently, it makes sense to identify a couple of promising fluorescent...

...Addgene was built on the concept of accessibility: making plasmids more accessible to scientists around the...our very first professional development workshop, “Make a Greater Impact Using Clear and Accessible Scientific... you can submit it with your manuscript and help make your paper accessible to even more people.  Whether...

...is the fact that Cas9 can be easily programmed to make a DNA double strand break (DSB) in the genome wherever...cleaning enzymes (Aprataxin, Tdp2) that function to make ends better substrates for ligation. Although we...Cas9-generated breaks is useful if the intent is to make a knockout in your gene of interest, as it is prone...

...20,000 genes, it is not possible to test each one and make sure it’s working. You can only do that screen with...system by screening for genes that, when turned on, make melanoma cells resistant to treatment with a BRAF...please ensure that you are equipped and authorized to make and use lentivirus. Also, note that due to the size...

...examples of the toolkit in action, see Cermak T, et al. 2017. Cermak T, et al. Plant Cell. 2017. PubMed ...functions and applications. We hope that these articles make it easier for you to find and use the plasmids you...that is easily separable from that of EGFP. This makes mCyRFP1 a powerful tool for use as a donor in FRET...

...and, most importantly, any connections the neurons make to other distant brain areas. It is desirable for...particles are relatively small (20 nm) in size which makes them capable of diffusing in the extracellular space...particles will infect neurons at different ratios, making these neurons fluoresce in different colors. An... Resources on the Addgene Blog Retrograde AAV: Making the journey from axon to nucleus Rabies and neuronal...

...end, you’ll have a better understanding of what makes polyclonal antibodies unique and what experiments...together, all the B cell clones in the immune system make different isotypes of antibodies that recognize ...other antibody formats. This low barrier to entry makes polyclonal antibodies a common reagent in many labs...

...and share some thoughts from the community that makes us… us. “The Addgene not-for-profit is an amazing...resource. I really appreciate the work you do to make sharing reagents and protocols so easy!” — Nina ...people knew how to access Addgene, and that we were making it possible for scientists to both deposit and ...

...one light chain. When assembled, the four chains make a Y-shaped protein that can be divided into the ...use Fab fragments as primary antibodies, you can make your own antibody fragments as needed.  In contrast...format. Reporter-conjugated anti-IgG Fab fragments make good secondaries for cases where you need to pre-form...

...Palladian Partners. “I was lucky to have been able to make that transition really smoothly,” says Susan. After... a very painful process. And certain stories can make you feel like ‘I’m a terrible writer. I have never...science communicators. “The power of a story is to make a human connection with a reader, whether it’s a...

...summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you...inhibition is spatially and temporally ubiquitous, making them a rather blunt instrument for controlling ...al, Nat Chem Biol 2018. PubMed PMID:29483642. Making protein-protein interaction studies easier with...

...scientists starting their own labs get excited about making their first deposit to Addgene. We are also ...cutting edge practices and benefits and these are making a difference. We serve scientists around the globe...about being able to do good in the world. We are making an impact.  {% video_player "embed_player" overrideable...

... always excited when depositors find new ways to make this workhorse protein even more useful! From FPs...retain normal neuronal function, this system may make optogenetics in specific cell populations even easier...applications, including optogenetics, and they should make it much easier to conduct functional studies in ...

...plasmid such as pX330. This cloning strategy not only makes it easy to create a single gRNA-expressing plasmid...clone up to 7 gRNAs into one destination vector, making multiplexing easy.  Disadvantages of Golden Gate...TAL effector-based constructs for DNA targeting. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt...

...biological. Even the most high-fidelity polymerases can make mistakes and lead to the accumulation of errors ...ensure the entire team is using the same rubric and making the same decisions when analyzing the wide variety...the QC team leaves a note on the plasmid page to make requesting scientists aware of the identified discrepancy...

...precipitates that permanently stain the membrane, making that section of the blot unusable for reprobing... if the mild is not effective. You may choose to make your own stripping solution using a recipe like ... Assess the stripping Now it’s time to check to make sure the antibodies from your first round of detection...

...it also helps you to hone your writing skills and make a name for yourself. Pick a topic that is close ...’t you record a podcast or put some effort into making a video? Use your unique talent to communicate ...Since then, I have tried many things: photography, making videos, presenting in pubs and at science festivals...

...crucial if we want to make sure that everyone can be informed and empowered to make their own decisions ...posts, lesson plans, or information briefs for policymakers. The rapidly changing landscape of genetics ...

...Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J,...plasmids from this publication at Addgene. 4. Makarova KS, Wolf YI, Alkhnbashi OS, Costa F, Shah SA, ...

...Requiring an additional plasmid for transfection can make these optimization experiments a bit more challenging...experimental volumes accordingly. While you may need to make some final adjustments, this is generally a good...additional resources. References Kumar M, Keller B, Makalou N, Sutton RE. Systematic determination of the packaging...

...vehicles for genetic information. Therefore they make excellent tools for genetic engineering. There are...vectors when used in experiments. Retroviral vectors make up a large toolbox used by researchers for (among...shRNA or miRNA. The ever expanding CRISPR field makes wide use of retroviral delivery systems, especially...

...spatiotemporal control or an “on/off switch.” This makes them great tools for providing a global overview... of the bait protein will be biotinylated. This makes it’s impossible to distinguish proteins that directly... proteins. This extended incubation period could make it difficult to detect proteins with short half-...

...proteins had to be re-engineered for each gene, making wide-scale gene activation seem next to impossible...activation compared to other CRISPR activators, making it a popular method for gene activation experiments...design (SunTag). This streamlines its delivery, making it a common choice for CRISPR activation. Newer...

...meeting in person is often much more productive and makes it easier to build relationships. Nonetheless, the...responsibilities Being a leader in an organization, you need to make sure the organization speaks in “one voice” whilst...such as leadership skills can only assist you and make you more qualified for future career opportunities...

... who keep changing their minds and have trouble making decisions. In my work as a thesis coach for graduate...be reviewed, bring them to the meeting. This will make it easy for your supervisor to support you. 3) Explain...articles on our blog Read Joanne Kamen's Post on What Makes a Good Mentor Resources on the Addgene Wesbite ...

...capacity and stable expression of lentiviral vectors make them an attractive option. However, safety requirements...has psuedotypes. Psuedotyping is the process of making a hybrid AAV, with the genome from one serotype...Klingler, E., Kuhn, M., Farouni, R., Lagler, S., Kalamakis, G., Lischetti, U., Jabaudon, D., & Platt, R. ...

... any features of the library backbone that would make the library more difficult to work with. For example...bacteria can grow and compete in the same container, making it easier for a smaller, recombined plasmid to ... reference protocols There are many decisions to make when developing your amplification protocol! We ...

...m going to share what I’ve learned and hopefully make your decision a little easier if an industry postdoc... your goal is to enter academia, it is easier to make that transition from an academic setting—especially...company you choose. If publishing is important to you, make sure that your project will not involve proprietary...

...Knockout/Knock-in plasmids Gene targeting methods also make it possible to insert, or knock-in, any gene, tag...instead want to create a conditional knockout. To make a conditional knockout, researchers often use the...floxed alleles. Although CRISPR is very good at making knockouts, knocking in large sections of DNA can...

...Addgene is accelerating this research by helping to make new cancer resources easily accessible to the research...found across all TCGA tumor samples is represented, making the results of any such study as clinically relevant...Addgene's cancer collection pages Scientists are making progress. New resources and technologies become...

...School of Medicine. In the Lindberg Lab we often make cell lines that overexpress genes of interest; more...Parents of infants will have no problem with this!) Make sure your colonies are well separated. For this,...taking both early- and late-arising clones, and making sure we always maintain a backup replicates of ...

...system), and the third post will describe how you can make your own mutant flies. Find Drosophila Resources...bristly hairs. Females do not have sex combs. To make genetic crosses properly, it is important to isolate...emerging at night. An incubator with a light cycle makes for very predictable work! When flies first emerge...

...the localization of your protein of interest is to make it detectable. Visualization can be achieved using... and their potential overlap, it is essential to make sure that the selected FPs do not influence each...your next localization experiment and invite you to make any fluorescent protein constructs you create in...

...usurped old methods or complemented them in ways that make them much easier. A special class of FPs, the FP...functionality with a single gene Although it is possible to make a timer system composed of two separate FPs (one... This one protein method simplifies cloning and makes your experimental setup less cumbersome. 3. Portability...

...that commercially available cloning kits can often make the process much more streamlined, because they ...the reaction in favor of the desired assembly to make the process close to 100% efficient. Gateway cloning...Multiple inserts: There are some cloning methods that make working with multiple inserts a breeze. Among them...