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TypeBlog PostPublishedJune 29, 2017, 1:06 p.m....improving the maps further after this initial launch so stay tuned for more updates. Overview of SnapGene powered...of common plasmid features such as antibiotic resistance genes, selectable markers, promoters, origins...cloning. With this more functional display, you can start thinking about your next cloning experiment early...map, you also have the ability to to download the static image file (PNG, see above) for your own reference...hosted here retain all the features shown in the static maps discussed above, but also have interactive...For example, if you hover over the ampicillin resistance gene, you’ll find the full name of the gene, ...direction (if relevant), and types. Types listed are standard GenBank feature types. Clicking on a particular...
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Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2
TypeBlog PostPublishedMay 26, 2020, 1:15 p.m....Piggybac transposon to stably integrate a reporter gene that contains a puromycin resistance gene followed by...prevent alternative start site initiation, the lab removed the native methionine start site. This first ...CRISPR reporters in the near future. SRIRACCHA (stable, but reversible, integrated reporter for assaying...: A donor plasmid that contains a puromycin resistance gene linked to an in-frame H2B-GFP. This visual...Pruett-Miller SM, Blanc V, Davidson NO, Madison BB (2017) A stable but reversible integrated surrogate reporter for... -
Viral Vectors 101: The AAV Retrograde Serotype
TypeBlog PostPublishedApril 27, 2023, 1:15 p.m....neurons are physically connected, we can begin to understand how the brain works. It’s kind of like “the head...in a powerful way. Using brain structure to understand brain function So far scientists have identified...itself across these regions is important for understanding how the brain functions. The basis for these...neurons are physically connected we can begin to understand how the brain works. There are two methods of... spinal cord (Wang, 2018, Metcalfe, 2022) and substantia nigra (Tervo, 2016). Discrepancies in transduction...such as those in the spinal cord. Now that you understand this new tool, go forth and trace some neurons... Additional Resources on the Addgene Blog Get Started with AAV Learn more about Lentivirus Find out More... -
How to Make Friends and Meet People at a Scientific Conference
TypeBlog PostPublishedAug. 7, 2014, 1:58 p.m....same phosphate of your favorite kinase is a good start. Check out Joanne's Reddit AMA Perhaps you think...interests. I favor small conferences where the attendees stay onsite and eat together because scientists have ...scientific terms when socializing at a conference, but start with a more general intro until you know if the ...problems. During the conference The conference may start in the airport. If you see someone traveling with...and put your phone away (did I say that already). Start with, “Is this seat taken? Can I join you?” Next...follow LinkedIn to see when your connections post. Stay in touch by liking, commenting and re-sharing their...connections when they get a new paper published. Stay in touch enough to plan to meet up at the next conference... -
Celebrating 15 Years of Scientific Sharing
TypeBlog PostPublishedJan. 14, 2019, 2:08 p.m....serve scientists around the globe and our own staff is starting to reflect this diversity, remarkably so for...We are delighted at the early outcomes and the ecstatic reception by scientists. The success of these...own to share reagents. I love the way scientists starting their own labs get excited about making their ...Addgene. We are also lauded by another set of stakeholders - the tech transfer community. In a recent law...love working with local organizations like Just-a-Start, the Northeastern co-op program, and with other ... Turnover at Addgene is about half of what is standard in most sectors. The Addgenies support a high ... -
Human Germline Editing Using CRISPR
TypeBlog PostPublishedAug. 10, 2017, 2:19 p.m....paper bring up? We’ll sift through the paper to understand what Shoukhrat Mitalipov and his colleagues have...from this work. Experiments & major findings Let’s start with the paper itself: Ma et al. were interested... the edited embryos proceeded to the blastocyst stage at similar rates to untreated controls. Ma et al...results in fewer eggs. However, the potentially staggering costs of such a treatment on top of the expense...Since the embryos produced were not being used to establish a pregnancy, the researchers were able to examine... some uncertainty. As biologists equipped to understand the results of this paper, we should be willing... paper represents a huge advancement in our understanding of how CRISPR will perform in embryos but also... -
CRISPRainbow and Genome Visualization
TypeBlog PostPublishedFeb. 28, 2017, 3:30 p.m....with physical map distances of 2 and 75 Mbp, with the calculated fluorescent distances correlating with... was developed in the lab of Addgene depositor Stanley Qi and used in Chen et al. to track telomere dynamics...relationship of the two genes over time, at a range of distances from 2-75 Mbp. Labeling multiple, intrachromosomal...with the previously established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that...degree of chromatin compaction even for this small distance! To the authors’ knowledge, this work represents...gRNA will permit labeling, but a newly expressed standard length gRNA would “switch” the system and induce... -
Antibodies 101: Fab Fragments
TypeBlog PostPublishedSept. 5, 2024, 1:15 p.m....chains and two identical light chains) each with a constant region and a variable region. The chains are covalently...protein that can be divided into the Fragment Crystallizable region (or the “Fc”) and two Fragment Antigen... of the antibody and comprises portions of the constant regions of the two heavy chains. In the lab, this...comprise the entire light chain and a variable and constant region of the heavy chain. As the name implies...labeled. V = Variable domain. L = Light chain. C = Constant domain. H = Heavy chain. Created with biorender.com...in super-resolution microscopy by reducing the distance between the target and reporter. They have fewer...tissue penetration. However, they tend to be less stable and have lower binding affinity than some of the... -
Working with Nuclear Receptors
TypeBlog PostPublishedJuly 15, 2014, 4:03 p.m....and disease states, their circadian regulation, as well as in important physiological states such as adipogenesis...which then return information such as nutritional status. Within the animal kingdom, this information is..., one of our primary goals is to promote an understanding of how these processes are regulated at the ...central and peripheral organs because of their established roles as molecular sensors and governors of endocrine-hormone...that interact directly with NRs) exist in steady-state complexes with secondary coregulator (or co-coregulator...whole complex. For example, the phosphorylation status of SRC-3 defines its association with other members... -
Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation
TypeBlog PostPublishedDec. 14, 2021, 2:15 p.m.... whether a certain methylation state causes a condition. To understand mechanisms and, eventually, develop...’s dCas9… with no modifications). Ready to get started? Figure 1: Break free from the ball ..., transcriptionally active dioxygenase When I started to work on building a CRISPR/dCas9-based tool for...expression and pLM-CMV-R-Cre. Selection for cells stably and highly expressing these constructs can be achieved...achieved by fluorescence-activated cell sorting, and stable gRNA expression can be achieved using lentiviral...backbones expressing blasticidin or puromycin resistance genes. It’s that simple! Good luck and happy... -
Viral Vectors 101: Virus Safety
TypeBlog PostPublishedJune 22, 2023, 1:15 p.m....infected by a lab-grown virus, it's important to understand the risks of the specific virus you’re working...with before you work with it. This will help you understand what safety precautions to take, and if something...ethanol (other surfaces). For AAVs, which are resistant to ethanol and isopropyl alcohol, 0.5% peracetic...waste, and of course, if you are working with a substance in a biosafety cabinet, it needs to be decontaminated... organization’s reporting mechanisms before you start an experiment, just in case something does happen...‘off’ day – you are both more likely to make a mistake and more likely to want to cut a few corners. ...dangerous things they work with, and that can understandably induce some nerves. Following safety procedures... -
Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux
TypeBlog PostPublishedSept. 13, 2016, 2:30 p.m....Dolan, F100Research, 2016). These events have also started progress towards solutions. The 2015 Boston FoR...landscape facing all scientists, both young and established, as they embark on or continue their careers ... upcoming implementation of the Federal Labor Standards Act (FLSA) which mandates that all workers across...graduate students who work as teaching or research assistants to join unions. The topic of unionization continues...problems facing postdocs and graduate students, and started to define solutions but this year, together with...of Howell Communications Philip Brenner, PhD - Assistant Professor of Sociology. University of Massachusetts...uses synthetic biology to create tools to better understand human disease. Outside the lab, he is interested... -
Illuminating Epigenetics with A FRET Based Biosensor
TypeBlog PostPublishedNov. 19, 2015, 3:30 p.m....and methylated states. Thus, this system is a reliable tool in reporting methylation status of these sites...the headlines, with sotires like the potential devastation of the palm oil industry through epigenetic effects...grandchildren; in this study,epigenetic changes were statistically significantly associated with reduced life-span...syndrome is characterized by low muscle tone, short stature, cognitive disabilities, problem behaviors, and...molecular biologic techniques to further the understanding of epigenetics; techniques include chromatin...Revolution: How Modern Biology is Rewriting Our Understanding of Genetics, Disease and Inheritance Amazon ... -
10 Basic tips for mammalian cell culture
TypeBlog PostPublishedJuly 12, 2018, 1:09 p.m....cells don't stay out too long While you’re preparing things, make sure your cells don’t stay out too long... on the hood for at least 15 minutes before you start any tissue culture work and make sure that clean...as well as that watch you’re wearing before you start. Also, avoid unnecessary talking because talking... the hood and can wipe them all down before you start. This will also help you avoid moving things in ...already be at 37 °C. Most labs have a water bath constantly at 37°C for this purpose.Learn about the effects...split very soon. There is power in numbers! When starting off with a new cell line, grow the cells, aliquot... -
Hot Plasmids: Summer 2025
TypeBlog PostPublishedAug. 28, 2025, 1:15 p.m....Here's what you'll find in this post: Bright and photostable yellow fluorescent protein mGold2 Viral delivery...synaptic connections with ATLAS Bright and photostable yellow fluorescent protein mGold2 By Mike Lacy... The mGold2 variants significantly enhanced photostability — 25-fold over mVenus and 4-fold over the previous...mGold2s, or mGold2t. Bottom: Quantification of photostability. Image reused from Lee & Lai et al. (2025) ... Lee, J., Lai, S., et al. (2025). Bright and photostable yellow fluorescent proteins for extended imaging... Figure 3: Using LARRYv2 libraries for state-fate analysis in ex vivo HSC expansion cultures....cultures. A) Experimental design. B) UMAP of cell states in ex vivo expansion cultures from days 7, 14, and 27... -
Antibodies 101: Flow Compensation
TypeBlog PostPublishedMay 16, 2024, 1:15 p.m.... For anyone just starting to use flow cytometry, it’s generally recommended to start compensating with... depending on your flow cytometer’s setup. For instance, BV510 and AmCyan have almost identical emission...needing to use your single-color controls as a starting point. Applying compensation to your experimental...your main population and singlets. Then, you can start to compensate on your experimental samples. Let'... what two markers your fluorescent markers are staining for. If a cell is positive for both markers, it...challenging! But we hope this guide can help you get started. Remember, you can always change your compensation... -
Of Myc and Men
TypeBlog PostPublishedJan. 19, 2023, 2:15 p.m....use in the lab? Take for instance, the commonly used Myc-tag. Who first started using it in protein tagging...larger than the substance P peptide, but still quite small. However, unlike the substance P tag, they found...experiment involved a peptide from the neuropeptide substance P. This protein had a highly specific antibody...meaning the tag could be quite small. However, substance P has an amide group added to its C-terminus post-translationally...future. These newer findings simply add to our understanding of how this antibody works and allow users to... -
PhD Applications After COVID
TypeBlog PostPublishedJan. 5, 2023, 2:15 p.m....applied to PhD programs in Biology around the United States in 2021, I know that this process only became even... even more complicated with COVID-19 and standards for hybrid, virtual, and in-person events change seemingly... I reach out to potential mentors?” “Where do I start with picking universities?” and “How does COVID-...prospective student of such-and-such university State how you found their lab (usually through the university...other. Before COVID-19, financial and logistical obstacles could make touring potential programs and universities...online meetings with current students, faculty, and staff have become a new way to allow you to get important...options are very handy in moments like that! For instance, during my application cycle, two schools on opposite... -
Addgene's Cancer Collection Pages Connect the Research Community
TypeBlog PostPublishedSept. 6, 2016, 2:30 p.m....cancer-related complications each year. Despite these grim statistics, new strategies for attacking cancer are yielding...the circulatory or lymph systems to spread, or metastasize. Many cancerous cells will die trying, as the...technologies become available almost daily, yet obstacles remain. One of the biggest challenges facing researchers...project continues to evolve rapidly and, with assistance from the research community, will soon expand...pathways with potential to confer anticancer drug resistance." Science signaling 7.357 (2014): ra121. PubMed... Resources National Cancer Institute: Cancer Statistics Page National Cancer Institute: Ras Pathway v2.0...Resources on the Addgene Blog Tackling Cancers' Drug Resistance with a New Screening Kit Learn How the Clon Tracer... -
Anatomy of a Plasmid Page at Addgene
TypeBlog PostPublishedFeb. 4, 2016, 3:30 p.m....plasmids that need extra stability, such as some of our viral vectors, in NEB Stable cells. Resource information...in the example above that the bread crumbs trail starts in the browse page, continues through the depositing...Addgene Sequences’). These sequences are provided in FASTA format (example) and can be analyzed using Addgene...Addgene’s single plasmids are sent as bacterial stabs at room temperature. Core of the plasmid page - ...the plasmid in your experiments: Backbone: The starting vector that the depositing lab modified to create...is shipped. Copy number (if known), antibiotic resistance, and optimal growth temperature are listed, as...