...England Biolabs and Labconscious. I was eager to learn about practices I could bring both to Addgene, and...impact as scientists. Many scientists are already leading the cultural change toward responsibility in our...polystyrene will likely end up in a landfill, but at least we can reduce the rate at which that happens and... it offsets the cost of regular waste services. Learn about how to start a glove recycling program at ...on culture. Encourage your workplace to become a leader in sustainability. Sign up for blog posts and...companies, and universities to be sustainability leaders. Take initiative to bring ideas to your workplace...

... sites in a promoter lead to higher transcriptional activation. Multiple nuclear localization signals ...a higher signal-to-noise ratio with a lower photobleaching rate. Given the power of SunTag, they attempted... ratio Less chance of phototoxicity Reduced photobleaching Simplified, long-term single-molecule tracking...SunTag plasmids have coveted “blue flames,” it’s clear that the system is broadly applicable across many...eBook Avoid Fluorescent Protein Oligomerization Learn about Small Molecule Fluorphores Resources on Addgene.org...

...basis of the Surveyor assay is that Surveyor nuclease cleaves double stranded DNA (dsDNA) at basepair mismatches...mismatches at the site of the edit. Using Surveyor nuclease followed by gel electrophoresis, these mismatches...mismatches are indicated by the presence of DNA cleavage products. The appearance of the 428 bp band indicates...I designed (listed above) worked well and I got clear bands using the conditions above. 2. Design your...product, you would still be able to identify the cleavage product. 3. The Surveyor kit had a lot of good...Before digesting the DNA duplexes with Surveyor nuclease, save some of the duplex and run it as an undigested...

...Loren Looger’s lab, Kaspar Podgorski and colleagues have now released iGluSnFR3 (Aggarwal et al. 2022). This... oxytocin, Daisuke Ino and Hiroshi Hibino and colleagues devised a “Modular fluorescence unit fused with...voltage sensor The François St-Pierre lab and colleagues have developed a new genetically-encoded voltage...limited. In a new preprint, Jared Toettcher and colleagues describe pYtags, a modular system for monitoring...dynamic range. With this in mind, Ed Boyden and colleagues set out to engineer a potassium ion biosensor... signal/noise ratio. Here, Takeharu Nagai and colleagues present an improved genetically encoded temperature...

...out-of-the-box ideas that might jump start your learning. 1. Create a peer mentoring group We don’t all...matter why, now is a great time to find a few colleagues to work together on career and life goals. For...scientist for show wrestling, founded a nonprofit, learned to play the ukelele...perhaps you get the idea....hope you all know by now that people get jobs at least 85% of the time because of relationships. It’s much...conversations. If you set yourself a goal of doing at least two informational interviews a month you will make...academia and not in academia is run by grant funding. Learning to write grants quickly, efficiently and effectively...

...start – it is the discussion after that will lead to learning. Listen to our podcast interview with Harvard... take informal minutes on what was covered and learned, especially noting action items that are identified...Harvard Medical School researcher Connie Cepko to learn about her mentoring style. Set goals with a mentor.... There are many advantages to being a Mentor: Learn other perspectives Develop coaching skills Encourage...Encourage your creative thinking Enhance leadership skills Support inclusion, diversity, and open communication...

...industry.” In his article On Leaving Academe, Terran Lane is asked by his colleagues and friends why he would...in the Boston area that demonstrate that 85% of leaving postdocs pursue a career outside the traditional... like this without sufficient data? Starting to learn about the diverse options and how to take advantage...there. Additional Resources on the Addgene Blog Learn about Possible Science Careers How to Negotiate ...for Scientists Series Resources at Addgene.org Learn About Careers at Addgene ...

...of your colleagues. The young scientists that are reading your peer-review comments will learn from them...delivering constructive critique can help your colleagues at the receiving end, if not for this specific...repositories will ensure that the results of your colleague’s hard work will be maintained and made available...comment differently if I would be speaking to a colleague in the office next door. So be kind, helpful, ...writing a review article Resources on Addgene.org Learn about depositing plasmids pre-publication Find molecular...

...of scientists from MIT tapped into this data to learn about trends in synthetic biology and DNA synthesis...database. Through a series of simulations and machine learning, they created a classifier that predicts whether...gene and a synthetic version of that gene was a clear predictor of whether the gene is synthetic or natural...applications in monitoring the accidental or intentional release of engineered organisms. “Is [the organism] engineered...Nielsen, Alec AK, and Christopher A. Voigt. "Deep learning to predict the lab-of-origin of engineered DNA...Addgene blog Read blog posts about synthetic biology Learn more about codon usage biases Resources on Addgene.org...

...on Precision Genome Editing with Programmable Nucleases. Check out #KSgenome on Twitter if you missed ...the CRISPR field forward. To do that, we need to learn much more about cellular DNA repair pathways. An...An emphasis on cellular repair post-CRISPR cleavage When I first looked at the conference agenda, I was...would open not with an introduction to CRISPR nucleases, but with a session on DNA repair. What stuck ...) Delivery The RNP delivery method consistently leads to high on-target editing and low off-target editing... tool supports over 250 genomes, and they just released a brand new CRISPOR manual to kick off 2018! "...

...rapid multi-part DNA assembly method - read on to learn how to use it in your research. The starting point...cloning used short sticky ends, LIC employed the exonuclease activity of T4 DNA polymerase to create longer...be present in the reaction mix, limiting the exonuclease activity to the first occurrence of that nucleotide... intermediates” through PCR and imprecise T4 exonuclease activity, overcoming the requirement for carefully...step, the addition of a single dNTP stops the exonuclease reaction. The PCR fragment and plasmid are then... DNA polymerase, Gibson assembly requires T5 exonuclease in combination with Phusion polymerase and DNA...

...Avoid star activity. Some endonucleases (for example BamHI) are capable of cleaving sequences which are similar...cloned the plasmid yourself or obtained it from a colleague down the hall, it is always a good idea to take...incubating your DNA with restriction enzymes which cleave the DNA molecules at specific sites and 2) running...manufacturers make High Fidelity versions of the endonucleases and/or supply custom buffers as means to avoid...is not just necessary but also an easy process. Please vist Addgene's resource for plasmid verification...

...back: David Gordon, Jeff Guo, Fengbo Zhou, Paddy O’Leary (UCSF). Our approach is to discover key human...FDA-approved). This is a big job, but under the leadership of Nevan Krogan at UCSF we’ve brought together...preclinical studies already. We are following up on these leads as we speak, with collaborators in Paris and New...affected in various ways by the pandemic, it is clear that we have to stick together (6 feet apart), to...shout out to Margaret Soucheray, Jeff Guo, Paddy O’Leary, Fengbo Zhou, Helene Foussard, and Dave Gordon)... plasmids for coronavirus or COVID-19 research, please email Addgene at [email protected] with your order...

...while the alcohol used in the staining process bleached another, Klebsiella pneumoniae. After realizing...safranin, as a counterstain to help visualize the bleached bacteria. The chemistry behind Gram’s crystal ...drop by drop onto the smear until the alcohol runs clear and immediately rinse. Flood the smear with the ...Rinse the slide in tap water until the runoff is clear, blot gently to dry, and then view under an oil ...peptidoglycan membrane is unable to retain the stain, leading to its decolorization. In contrast, counterstain...visualizing differentiations between bacteria, leading to safer medicine and more successful experiments...

...drug discovery, and synthetic biology. Read on to learn more about the five awardees and their promising...their use as food, infusions of L. taraxacifolia leaves are widely used to treat several diseases. Her ..., the most common type of stroke and the third leading cause of death and disability worldwide. An ischemic...ischemic stroke occurs when arteries leading to the brain become blocked, resulting in reduced blood flow...awards. Additional Resources on the Addgene Blog Learn More About the Addgene-Seeding Labs Plasmid Grant...Expression Systems Synthetic Biology Resources Learn about Global Plasmid Distribution ...

...game that would force my students to use concepts learned from previous lectures, articles and books. A game...board with them. My impression was that they had clearer ideas about gene therapy in the following lessons...perhaps the ideas spurred by this “gamification” will lead to new cures for disease someday! Accessing and ...modify BUILD YOUR PLASMID or generate games like it, please freely share your projects with us! This exchange...want to receive our modifiable powerpoint file, please feel free to contact us at: [email protected] tools for teaching developmental biology Learn how BioBuilder can help teach students about Synthetic...

...which cleaves the synaptic protein VAMP2, a protein essential for synaptic transmission. Cleavage of VAMP2...ever like to write about a recent plasmid deposit please sign up here.      Listen to this episode of the... it inactive, thus inhibiting neurotransmitter release. How is this light-mediated? The researchers split...split botulinum neurotoxin halves to enable VAMP2 cleavage, and suppresses neuronal transmission within minutes...differentiation. Furthermore, TALEN and ZFN require custom nucleases for each gene of interest. CRISPR-Cas9 tools have...

...polymerases (Pol μ and Pol λ), nucleases (Artemis), and structure-specific end cleaning enzymes (Aprataxin, Tdp2...Cas9 protein doesn’t immediately release from the broken end after cleavage, which may interfere with loading... the products of accurate repair are easily re-cleaved while indel products aren’t, so repeated cycles...latter products. As noted above, a single cycle of cleavage and accurate repair should take less than an hour...

...activate essentially any gene,” she says. The technical leap from mutating and deactivating a gene or genes to...published in Nature in December, Konermann and her colleagues in Feng Zhang’s lab describe just such a system...transcription activation processes. Konermann and her colleagues demonstrated the new system by screening for ...in those 15,977 human genes. Weissman and his colleagues demonstrated in a paper in Cell that they could...which guide RNA we use. What we have to do now is learn more about the rules.” He said that there is no ... These libraries are lentiviral in nature, so please ensure that you are equipped and authorized to ...

...specific. Once inside the host cell, the virus unleashes its cargo – its own genome.   Fig. 1:...access to the host genome requires breakdown of the nuclear envelope, which occurs during mitosis. The most...and dividing cells. They accomplish this through nuclear localization signals which can traffic their viral...PMCID: PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human immunodeficiency virus type 1 preintegration...PMC1368960. Additional Resources on the Addgene Blog Learn More about Lentivirus Listen to Connie Cepko Discuss...Resources on Addgene.org Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral ...