...can be engineered to be cell type specific. Once inside the host cell, the virus unleashes its cargo – ...transduction efficiency and can effectively infect a wide range of cell types that are typically ‘difficult...viruses have a low immunogenicity which makes them ideal for applications with whole organisms. Finally, ...right approach for your experiment, you’ll need to decide exactly which type of virus to use. Types of viruses...genome into the host genome, setting up permanent residence. They primarily infect dividing cells, as their...signals which can traffic their viral particles inside the nucleus, without the need for envelope disassembly...clinical applications, viruses are true bench to bedside tools. Keep an eye out for our upcoming blogs on...

...infectivity): The use of the VSV-G envelope provides the widest tropism or range of cells a virus can infect...The widespread use of γ-retroviruses (gamma-retroviruses) in cancer and stem cell research has prompted...into the 293T packaging cell line. This system provides the greatest flexibility to pseudotype γ-retrovirus...encoded by the transfer plasmid, but instead are provided in trans by other plasmids or the packaging cell...terminal repeats; U3-R-U5 regions found on either side of a retroviral provirus (see below). Cloning capacity...Protocols: General Transfection The Addgene Retrovirus Guide  ...

...collection that grows on a daily basis, the lists below provide a snapshot of relevant resources and a good jumping...the gene you’re looking for on this list, please consider searching directly on our website. In-exhaustive... from a Set of Mitotic Chromosomes. Samwer M, Schneider MWG, Hoefler R, Schmalhorst PS, Jude JG, Zuber...DW Cell. 2017 Aug 24;170(5):956-972.e23 Screen identifies bromodomain protein ZMYND8 in chromatin recognition...receptor signaling. Zhao K, Wang W, Rando OJ, Xue Y, Swiderek K, Kuo A, Crabtree GR Cell. 1998 Nov 25. 95(5)...chromatin and the epigenome. If you have been considering some new techniques, the posts below may steer...

...Microscopy, University of Amsterdam. Before you decide which car you want to buy, it is worthwhile to ...tables with fluorescent protein properties and provides a quick way to compare fluorescent proteins, e.g... the protein of interest by a 2A self-cleaving peptide (Goedhart et al, 2010). An explanation of the assay...proteins (with the same reference protein) under identical conditions, a ranking of the practical brightness...practical brightness. Therefore, practical brightness provides a better picture of what can be expected in a ...theoretical brightness. Consequently, it makes sense to identify a couple of promising fluorescent proteins and...mimicking the future application. In figure 2 we provide an example of the comparison of the practical brightness...

...Viral Service. With their support we are able to provide faster access to a larger number of viral preps...Karpova’s lab from Janelia Research Campus have provided valuable insight into AAV production. Finally...advice and encouragement. Why virus? Why did we decide to take on this project? Addgene’s mission is to...plasmids in a ready-to-use viral format. Having identified this need in the community, we approached it ...control, generates  educational materials, and provides viral vector technical service (details about ...expertise in answering questions on virus and helping provide educational resources on this collection. With ... we could ship internationally and be ready to provide extensive logistics support for export and shipping...

...modification of multiple genes is often necessary. Guide RNAs (gRNAs) are commonly packaged in 400-500 bp...terminator. These relatively large cassettes (considering the gRNA itself is ~100 bases) limit the number...cleaved precisely as predicted with no addition of nucleotides to the 5’ spacers. The 3’ ends were modified ...tRNA-gRNA-tRNA constructs. gRNAs with any 5’ leading nucleotide could be processed, a distinct advantage over...over other methods that favor certain nucleotides at the 5’ end. PTG construct schematics. Scissors indicate...frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct loci. They also examined...this publication at Addgene Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing...

...transformed the palette of experiments possible in a wide range of organisms and cell lines. In C. elegans... on repair of a conditional-lethal mutation to identify edits with minimal screening effort. Selecting...504 restriction digests of PCR products used to identify a single 2xFLAG knock-in by direct screening. ...several aspects of editing. First, they devised guidelines for designing single-stranded repair templates...from a single injected animal.  PAM sequence considerations For changes 5’ to the PAM, sequences corresponding...non-homologous sequence or a series of single-nucleotide substitutions along a 1.5 kb tract, all without...CRISPR in C. elegans The Meyer and Mello labs also identified several experimental set ups that could improve...

...sequencing. Sequencing the 16S rRNA reveals the identity of the organisms present while sequencing of all...gives clues into what the microbes are doing. But aside from sequencing, scientists can also study the microbiome...limited and many microbes are difficult to culture outside of their natural environment. Thus, the Harris ...tested this system in vitro and in vivo. They identified cells from 19 genera across four phyla that have...conjugation in vivo studies. However, they only identified seven genera in in vitro experiments, possibly...Clostridium difficile. As mRNA expression cannot be identified with DNA-based qPCR methods, the ability to detect...Monitoring microbial population dynamics in the gut can provide insight into how the microbiome responds to infection...

... manuscript whose whole purpose is to provide detailed evidence of knowledge new to mankind. And you probably...logical to a layperson. When you’re communicating ideas that you think are clearly connected, step back ...overwhelming your audience - like the professor who once decided that to teach high school students PCR, they first...links to pay attention to. If you’re watching a video, you now have moving images, noises and music, spoken...olds is unlikely to be able to follow a YouTube video on PCR, while a senior lab scientist is likely to...distraction, such as pausing or turning off the slide deck (or reminding them to put their phones away...

...across the globe.  “Read the Addgene guide: https://www.addgene.org/guides/crispr/” — SirHamhands, responding... scientists. She, Kenneth Fan, and Benjie Chen decided to start an open science nonprofit that would facilitate...navigate. This meant we could grow accessibility alongside our repository.  “I’m sure you hear it all the...created are for the community. More than 100 labs worldwide use our plasmids. I hope they contribute to great...we realized we could use the same principles to provide recombinant antibodies, another plasmid-based product...including our new Antibodies 101 eBook, the Antibody Guide, and two antibody animations.    A blast ...

...in cells? This question is largely unanswered. Ideally, fluorescent proteins should act as inert modules...understand how well fluorescent proteins approach this ideal situation, cellular assays have been proposed. Early...homodimerization tendency. This assay employs a peptide derived from cytochrome p450, CytERM, to direct... looking cells can be counted. This assay will provide information on the dimerization tendency in living... interest will be problem-free. In figure 1 we provide an example of the OSER assay with a number of fluorescent...mScarlet-I shows correct ER labeling, which is good evidence that mScarlet-I acts as a monomer in the cell ...preprint, or documenting it in some other way. Besides the data, new plasmids and/or cells or organisms...

...LC3 doesn’t provide information about the contents of the autophagosome nor does it provide information...an autophagy inhibitor. While this method does provide a quantitative measure of autophagy, it only measures...studying autophagy. This method relies on the identification of autophagic structures based on morphology...morphology. Autophagosomes are relatively easy to identify: double-membraned structures containing undigested...with the vacuole or a lysosome are trickier to identify because their contents can be at various stages...Protein Plasmids and Resources Fluorescent Protein Guide: Biosensors Keima, another fluorescent pH-biosensor...

...is planning or might need to plan for a career outside academia (that would be pretty much all scientists...usually visible to you (unless they’ve chosen to hide it). While most people use LinkedIn messaging, not... should be asking yourself if it’s really a good idea.  Pro tip: Don’t hit the connect button unless you...your network. If someone has connected with me, I consider them fair game for connecting to others.  Pro ...can type a full note with good format without accidentally sending before you’ve had a chance to proof ...including and especially the hiring manager. This is considered intrusive and it’s still connecting with a stranger... people’s news and updates. Share your own good ideas, events and favorite resources. Respect the network...

...train our young scientists for futures both inside and outside of academic research (4). There are currently...all bad, however - there are some signs that the tides may be turning. In 2013, the NIH announced its first...finding solutions to some of the problems that were identified last year.  In particular, at the symposium we...few or too many at different levels? What does an ideal and productive workforce look like for the sector... Law School. She recently launched a survey “Identifying Career Pathways for PhDs in Science”, through...create tools to better understand human disease.  Outside the lab, he is interested in communicating science...

...part of her larger tutorial series, A Beginner’s Guide to Writing Scientific Manuscripts. Any scientific... first time. The first time through, just get an idea of the techniques used in the research, but wait...with the literature in your field of interest. Set aside some time each day or week to at least scan the ...Google Scholar alerts. Based on keywords that you provide, Google Scholar will send you daily updates with...library as well as some integration with internet-wide literature searching. Update 5/6/2015:Here's another...below. Additionally, read more of the Beginner’s Guide to Writing Scientific Manuscripts series on Johnna's...@JLRoose.   Additional Resources: A Beginner’s Guide to Writing Scientific Manuscripts by Johnna Roose...

...overwhelmed. Jumbled thoughts such as “I have no idea how to write a grant!”, “I don’t have time for this... setbacks, and potential alternative routes, Provides opportunities to convince others why a project...the RFA due date in order for the administrative side of the grant to be established. Take time to read...sections in a NIH RFA, for example, and tips to consider while writing each section. Research challenge...matter. The more detail (within the page limits) provided in this section will convince the reviewers that... the application. Make sure to invest time and consider the reviewer's comments. Whether the proposal ...comments help gauge room for improvement, deliver outsider opinions, and reveal items to include for the ...

...Bimolecular Fluorescence Complementation (BiFC). It can provide a qualitative or quantitative readout of the interaction...GFP(11)). Some of the articles linked below only provide one fragment of the FP because they are used with...pairs with N173 derived from Cerulean or Venus Identification of new fluorescent protein fragments for bimolecular...Huang Yellow Venus pBiFC-VN173, pBiFC-VC155 Identification of new fluorescent protein fragments for bimolecular... toolkit from Benjamin Field’s lab. Special Considerations There are a few special exceptions to the usual...reversible complementation. TagBiFC (split HaloTag) provides maximum flexibility in color labeling with exogenous...constructs or certain cell environments). Also consider that a protein fusion may disrupt functionality...

...strong immunogenicity of AdVs is what makes them ideal candidates for applications in oncolysis and vaccination...co-infection with a helper adenovirus is required to provide the viral proteins in trans. The Gutless AdV have...plasmid construct, please see Addgene’s Adenoviral Guide webpage, and references 1, 2.*Pro-Tip*: once the...the correct recombinant pAdV plasmid is identified, re-transform into a bacterial strain not prone to recombination...standard method for purification of rAdV uses cesium chloride (CsCl) density gradients combined with ultracentrifugation...PubMed PMID: 17546019. 2. He TC. et al. A practical guide for using the AdEasy System. 3. Jager, Lorenz, et...Addgene.org Find Adenovirus Plasmids Adenovirus Guide Page  ...

...and Alexander A. Green. Zika background First identified in 1947 in Uganda, the Zika virus had received... Aedes mosquitoes (aegypti and albopictus) and, aided by international travel, the Zika virus is expected... until vaccine programs can be put in place to provide community protection. Toehold switches for pathogen... enabled gene circuit-based sensors to be used outside of the laboratory. The detection threshold for ...draw on as our assays grew more stringent, and it avoided any days of lost productivity waiting for follow-up...of the pathogen’s genome as unique barcodes to identify samples containing the virus. Of the 48 sensors...genotyping, as well as our electronic reader that provides portable quantification of diagnostic results....

... spot when an application is rushed (whether the idea isn’t properly researched / developed or the application...sometimes be tricky, as most of our research builds on ideas and findings from our own or other groups, but your... with specialists in different fields that can provide different insight into an area - not just working...engineering new applications to solve problems will provide originality to your work and possibly a new perspective... your network and your collaborators It’s a good idea to move around to different labs during your career...make it clear what the outcome of the work was. Ideally this would be results published in a paper, but...their grant submission panel. Some funding bodies provide opportunities to attend an overview of what they...

...missed with conventional FP imaging. PA-FPs are divided into 2 main categories - those with irreversible...requires a temperature below 30°C and this is not ideal for experiments in mammalian cells. Dendra2 is another...light both activates and inactivates Dronpa is not ideal, since this means Dronpa is quickly inactivated ...your experiment, there are several parameters to consider, such as brightness, photostability, pH stability...protein aggregation. Another important parameter to consider is the contrast ratio- a comparison of PA-FP brightness...intensity used to activate the PA-FP should also be considered. If the light intensity needed for activation...to preserve cellular structures (0.5-1% osmium tetroxide). These conditions destroy most PA-FPs, but, in...

...Is the conference you are attending on the larger side? At large conferences, chances are that several ...have additional questions. If you or your lab identifies a potential collaborator from the meeting, reach...If your poster material is nearing publication, consider uploading your work to a preprint server (and ...Perhaps some of your poster visitors had a great idea or interesting questions worth discussing with your...Reach out if you’d like to meet someone and set aside a time to meet. Some conferences may also have a... of the things we've mentioned above could be considered networking, but here are some more general tips...resume and prepare business cards. You might not consider being a student or postdoc a “business,” but having...

...goal was to enter industry since I was drawn to the idea of working as a team to develop therapies that could... of academics tend to view industry as “the dark side,” that you’ve sold out. You’ve given up your intellectual...ve discovered.) As someone who has now seen both sides of the postdoc experience, I’m going to share what...an industry postdoc is something that you’re considering. Like anything, there are pros and cons to postdocs...will hire a group of postdocs and then let them decide what they’d like to work on later. For instance...suit your needs better than others. Whether you decide to do an industry or an academic postdoc, you’ll... open positions at Addgene Check out our career videos ...

...treatment or prophylaxis of COVID-19. Researchers identified nanobodies that bind to coronavirus spike proteins... randomized with a defined set of different trinucleotides (corresponding to different amino acids) in...bind to the antigen. The Seeger lab can directly provide the three sybody libraries in the form of mRNA ...tags are used in a final enrichment step for identifying sybodies that bind strongly and specifically ...their protocol because it is less susceptible to identifying unspecific and low-affinity sybodies (Zimmermann...-30) of sybody candidates are sequenced for identification. All unique sybodies can be subcloned into ...demonstrated they could use the protocol to rapidly identify and isolate sybodies targeting the SARS-CoV-2 ...

...such a system is likely to introduce additional nucleotides flanking the insertion. If the DNA ends anneal...MMEJ may also introduce, substitute, or delete nucleotides in addition to the expected deletion, but the...breaks are necessary for knock-in: one on either side of the GFP-Puro cassette and one in between the ... last coding exon of your gene of interest. For ideal use, this gRNA should be cloned into a vector containing...cuts in a staggered pattern, it is thought to be ideal for HDR-independent knock-ins, but this possibility...Engineering Tools like Sleeping Beauty Mutase Find the Ideal CRISPR Software for Your Experiment Additional ...Addgene.org Learn All about CRISPR on Our CRISPR Guide Pages Find CRISPR Plasmids for Your Experiments ...

...-Cy7 and PE-Cy7; or BV421 and BV51. It is a good idea to look up new fluorescent markers before deciding...them, using a tool like BioLegend’s Fluorophore Guide Chart.  Pro tip! Some antibodies go by different...setup. For instance, BV510 and AmCyan have almost identical emission spectra but may be called BV510 on one...which way your population needs to be shifted, consider your perpendicular lines. In this case, the population...APC-Cy7 are both bleeding into each other, as is evident by the two diagonal populations in the plot. A ...applying very large compensation values (100+), consider what two markers your fluorescent markers are ...fluorescent markers can be challenging! But we hope this guide can help you get started. Remember, you can always...

...you reach for that silica spin column, stop to consider some ways to purify DNA without a kit. DNA purification... kits have advantages: they are convenient and provide uniform, consistent results. But they are also ...method described in Kim and Morrison is nearly identical to that of commercially available kits but instead...concentration of a chaotropic salt (guanidine hydrochloride for the plasmid purification protocol and guanidine... Image: Kim and Morrison, 2009. This method provides DNA yield and quality similar to that obtained.... Homemade buffers performed the same as those provided with the kit. Although not tested by the authors... Molecular Biology Reference Visit our protocol video library References Kim, Y., & Morrison, S.L. (2009...

...Working with AAV vectors in neuroscience Let’s consider my first challenge when I started working with...). However, this one set of experiments didn’t provide any information on other important factors such... science initiatives, like Addgene’s Data Hub, provide an avenue to more easily solve these frustrating...every researcher experiences and that it often provides incredibly valuable insight into one’s work. As...different methods and vectors, and use this data to guide them in their own experiments. By providing DOIs...highlights the dearth of AAV vector information provided in openly accessible forms like publications. ...Sommer, J. M., Pierce, G. F., & Qu, G. (2005). Identification of factors that contribute to recombinant AAV2...

...biological disciplines and scientists employ GFP for a wide number of functions, including: tagging genes for... in the wildtype protein and facilitating its widespread use in research. Many other mutations have since... that many fluorescent proteins found on the red side of the spectrum are not GFP derivatives, but are...folding, maturation, and fluorescence. Biosensors: A wide array of GFP-based fluorescent biosensors has been...plasmids often include GFP as a marker to help identify which cells have successfully taken up the plasmid...applications for GFP. It has also been used to identify particular cell populations in drug screens, to...Plasmids 101 Articles Addgene's Fluorescent Protein Guide Note: A. Max Juchheim contributed to the writing...

...hypoxanthine (shown) is the nucleobase component of the nucleoside inosine. Created with BioRender.com. Inosine...act on newly-deaminated inosines. Pro tip! The nucleoside inosine consists of the nucleobase hypoxanthine...transversion editor AYBEv3. This tool still retained considerable A → G editing activity at most sites, in addition...initial editor AXBE, for A → X base editor (X = any nucleotide). To create a more specific A → C base editor...be useful for saturation mutagenesis, and they provide a strong proof of concept for further development...efficiency, purity, and indel rate of each tool vary considerably. And there’s so much more exciting protein engineering...base editing Get a broad overview with our CRISPR Guide Check out our CRISPR 101 ebook ...

...site, and titer. Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult... bias: Addgene uses a prioritization process to decide which tools we make available. This prioritization...community. It does, however, necessitate us first identifying broadly useful vectors, a process that isn’t ...information from requestors and depositors, and identifying trends in our own repository data, giving us ...viral vector educational resources, from eBooks to guides to protocols, which was why I was so excited to...2023; Haery et al., 2019; McDade et al., 2016), video, educational resources, and collaborations, and ...Swanson, L. E., & Fan, M. (2016). Practical Considerations for Using Pooled Lentiviral CRISPR Libraries...

...around $45,000, but as I’ve discussed elsewhere (slides here) there are many postdocs paid at much lower...must be met. In the Department of Labor’s general guide to the effect on higher education, you can see that...and Brandeis universities have stated that they consider postdocs paid on stipends to be FLSA exempt, and...institutions; and that institutions which can more easily provide bridge funding to absorb the cost will encounter...jobs that the majority of postdocs end up getting outside academia. There is also a valuable lesson for the...level they will be raised to on Dec 1st - across a wide consensus of academic stakeholders. There was ample...going away, and it is becoming ever clearer that outside scrutiny of science and the academic system is ...

...letters, while higher-level jobs are more likely to consider cover letters an important part of the application... content team, where writing is a core skill, considers cover letters to be very important. On the other...part of the job application process, it's a good idea to talk to people in the field to understand what...cover letter for? A cover letter is a place to provide helpful context about your candidacy, beyond what...make it easy to master ddPCR.  Finally, it’s a good idea to state why you’re applying to the specific company...unofficial poll, hiring managers also said that they considered cover letters to be more important for higher-level...Additional Resources Hamilton College Cover Letter Guide How to Write a Cover Letter Blog Post from Hubspot...

...invariably inconclusive and unreliable. Proper controls provide the constant variables that enable the correct ...functionality of your experimental system and help identify opportunities for troubleshooting or optimization...experiment, untreated cells should be included as these provide the baseline/standard against which other samples...correct interpretation of the results because it provides an important reference point when analyzing the... but the papers cited in the reference section provide more in depth information. Technical replicates... time consuming and ultimately more important. Ideally, each biological replicate should use fresh media...plasmids, be performed on different days, etc, but outside constraints may impose some limitations and you...

... of multiple animals on different occasions via video, generating huge amounts of data. When published...experiment would require access to each individual video. To PLOS’s credit, the organization subsequently...subsequently clarified what the policy intent was and provided an FAQ list to further explain the policy. The fact...variation is calculated and placed in the legend. Widely reported difficulties in reproducing experimental...the author or their host institution expected to provide data in formats exported to common file types? ...could be mitigated by on-line services such as SlideShare, Figshare and Dataverse. Just as Addgene helps...than just a single projection of it, could allow idea generation and extension, based on existing, published...

...with them. If you decided to make your own competent cells using the protocols provided above, then you ...cells are available online. A few examples are provided here for reference: OpenWetWare Preparing chemically...phase, then wash and resuspend the cells in the provided buffers. The competent cells are then aliquoted...Stbl3TM. How to transform your plasmid DNA If you decided to use commercial competent cells, then you're ...transforming your cells? Check out our transformation guide.   ...

...mutations found in leukemia, the Lowe lab was able to identify a set of mutations that made cancer cells more...mouse line. Over the years the fly has become an ideal model organism to study an array of topics including... to create transgenic animals. Zebrafish are an ideal model organisms for embryonic development studies... genes have an ortholog in C. elegans making it ideal to study basic molecular biology processes. C. elegans...quiescent and proliferative pancreatic β-cells identified by HOTcre mediated labeling." Proceedings of ...Menezes, Regina, et al. "From the baker to the bedside: yeast models of Parkinson's disease." Microbial...Additional resources on the Addgene blog Read our quick guide to Drosophila Download the fluorescent proteins ...

...loading control: the intensity of each POI band is divided by the intensity of the loading control in the ...problems for proteins of very different abundance, consider the following example gel. Figure 5: An...intensity axis. But the loading control points are outside of the linear detection range. As a result, they... problematic for computational quantification. Outside the linear detection range, band intensity measurements...experimental conditions. Otherwise, you might want to consider a different loading control. Assumption #2: The...final blot. Quantifying the entire lane of protein provides a loading control that can be used for normalization...The necessity of and strategies for improving confidence in the accuracy of western blots. Expert Review...

...Addgene’s catalog is a new class of sensors for neuropeptides and neurotransmitters. Motivated to develop ...GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost A to Z! - from ATP...vectors (here and here).  Figure 3: JEDI-2P was identified from a high-throughput screen, selected for increased...existing sensors. Using genome mining and structure-guided engineering of potassium binding proteins, they...point for further engineering. Figure 5: Structure-guided design and genome mining both led to new sensors...Fluorescent Potassium Indicators through Structure-Guided and Genome Mining Strategies. ACS Sens. 2022 May...protein plasmids and resources Fluorescent protein guide: Biosensors Collection ...

...New ideas and approaches are encouraged in science. It isn’t surprising that new ideas are also popping...before you even receive the product? How can we consider the environmental impact of those processes? Who...might sound risky in terms of contamination, but consider flow cytometers. These fluidic machines handle...through an EPA-approved process and no longer considered hazardous. Then, they use this material to produce...plastic lumber and distribute it to industrial and residential markets. Rethinking our approach to lab waste...

...Accessibility Guidelines (WCAG). These changes mean our content is both available and accessible to a wider community... Over 150,000 plasmids empowering researchers worldwide. Countless breakthroughs enabled by shared resources...part of the story. This year, we made significant strides in making science more accessible and reliable....community. As we enter 2025, your feedback continues to guide our innovation.  In response to a demand for more...rapidly, and with it, the needs of researchers worldwide. Through our growing community on Bluesky, our...

...pathway. “The idea is that if you introduce them into cells one at a time, you could identify those that ...apoptosis. For each of these 17 pathways, they identified a set of one to three mutant complementary DNAs... Wood says. “We want to make these available as widely as possible.” The researchers have plans to expand...References 1. Martz, Colin A., et al. "Systematic identification of signaling pathways with potential to confer...Lineage Tracing Read How CRISPR Can Be Used in Genome-Wide Screening Browse 28 Hot Plasmid Technologies from...

...How to put the wider scientific community at the service of COVID-19 research? We decided to give it a ...internationally COVID-19 Resources Canada is a database that provides information about reagents, funding and volunteer... the information is applicable to researchers worldwide. “COVID-19 has spread incredibly fast and without...can advance COVID-19 research to hopefully help guide policies and ultimately treatments in a timely fashion...Read about COVID-19 research on the blog Visit our guide for external collaborations Resources on Addgene.org...

...an introduction to plasmids. Plasmids 101 will provide you with an overview of general molecular biology.... Our mission is to curate a one-stop reference guide for plasmids, so that you can spend less time researching... such as yeast and plants. In nature, plasmids provide one or more functional benefits to the host such... biologists to broadly utilize plasmids across a wide range of applications. Some common plasmid types...scientists can track their location or quantity inside a cell Plasmids 101: Green Fluorescent Protein...references, and how to maintain your plasmid stocks. The guide also contains multiple protocols and troubleshooting...

... at the Addgene Science Career Guide eBook Addgene Science Career Guide eBook.   Resources Resources... a master? Here are a few questions to help you decide if a master’s might be for you. 1. Are you interested...path one you could see yourself taking? If so, consider setting up an informational interview with them...your interests and career goals. You might still decide to earn a PhD, but if I had to do grad school over...eBooks from Addgene, including the Science Career Guide\  ...

...but overwhelmed; I had no idea where to start. Thankfully, my managers set aside time for me to complete...the lab! I got to assist production on a protocol video. Don't worry - no pipettes were harmed in the making...allowed me to succeed. As a team, we talked through slide design, discussion items, big takeaways, and timing...depositor welcome kits, note taking for protocol video production, and keeping inventory of our merchandise...talk to. This six month experience made me more confident in myself, my abilities, and my future - something...

...middle of the targeted region, as shown below.  To identify optimal nickase designs, IDT scientists Mollie...insert an EcoRI site using a single-stranded oligonucleotide (ssODN) donor. Using a D10A nickase to insert...nickase outperformed both WT Cas9 sites tested by a wide margin, exhibiting >20% repair efficiency. Clearly... is especially useful if there are no available guides close to the desired mutation site. Quick tips...small insertions or tags (ssODN; IDT Ultramer® Oligonucleotides) Use 100 nt homology arms for large insertions... Find CRISPR Nickase Plasmids Browse the CRISPR Guide ...

...every project we undertake and every service we provide is tested against fulfilling our mission. “Addgene...continuously improve all the extra services we provide like quality control, curation, technical service...starting new initiatives as opposed to fundraising.   Aside from enabling us to make all decisions with the ... keeping excellent records and maintaining confidentiality where it is appropriate. This grows our relationship...Addgene increases productivity by providing world-wide access to curated, high-quality data and reagents...processes, distribution, and customer service) Addgene provides unmatched customer service that is consistently...

...pour over the sides.  These tiny gels and tiny lanes required less agar, less ethidium bromide, less stop...discards according to the Cambridge, Massachusetts guidelines and wear an extra sweater so we don’t have to...smaller gel apparatus until I settled on using glass slides. With just the right amount of agarose, I could...could pour a very thin gel on a large glass slide using only surface tension.  It's a bit tricky to pour ...plastic and solder this thing together.  One of the downsides—there was no cover so you had to be careful not...

...in 2012. Using structural modeling and structure-guided direct evolution, Nathan Shaner and colleagues ...respectively. mNeonGreen can be used as a fusion tag for a wide range of applications from traditional imaging to...resolution imaging. mNeonGreen also has little sequence identity to other common fluorescent proteins and therefore...The team that developed mTFP1 further sought to identify why this protein was blue-shifted compared to ...blue-shifting more than others. During this process, they identified mWasabi, a bright, monomeric green fluorescent...variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and ...