... plasmid via sequencing or restriction digest. Chances are, you’ll even have DNA left over for other applications... the culture medium. Improve the culture media Changing to a richer media or adding supplements can improve... low copy plasmids or slower growing strains, a change in media may be advantageous. Reduce the antibiotic...oxygen for optimal growth, as insufficient gas exchange will prevent your cultures from reaching the desired...shaker speed is fast enough for sufficient gas exchange in the overnight culture. Generally, large cultures...

...conformational change to bring two FPs on a single protein closer together or further apart. Changes in distance...dimers can form in the absence of environmental changes that the biosensor is designed to detect resulting...reporters for your studies and greatly lower your chances of finding artifactual results. Many thanks to...14581454. PubMed Central PMCID: PMC2173526. 2. Zacharias, D.A., et al., Partitioning of lipid-modified ...

...enable plant science in general. We are funded by a charity and since we are making all the tools for plant...fluorescent proteins. Visualizing fluorescence can be challenging in plants because of the auto-fluorescence of...U.K. organization called Athena SWAN that is a charter for advancing women in science. Universities sign... been a lot of engagement with the Athena SWAN Charter and it has really strengthened the network of female...tell them to work in plant science. I think their chances of doing something amazing will be equally high...

...coordinating the annual Math & Science Night, a chance for parents and other members of the community ...district that has a lot of parents in STEM careers, chances are they’re wondering how they could contribute...etc. Though I talked about CRISPR’s molecular mechanism, I was careful to keep it relatively simple. I... key concepts (Fig. 2). One thing I will likely change for next time is giving fewer examples and think...much in the making. One of the exciting, and challenging, aspects of talking to 8th graders was the broad...

...plants produce lots of seeds. More seeds mean more chances to recover a transformant. Use young plants: If...flowers to Agrobacterium. More seeds mean more chances to recover a transformant. Use lots of plants: ...per construct I transform. More seeds mean more chances to recover a transformant. Remove mature siliques...Agrobacterium all over your gloves, so be sure to change gloves between constructs if you are transforming...This won’t hurt them, and it will increase the chances of tDNA integration. Selecting Arabidopsis transformants...

...you select? In this blog, we’ll break down the mechanism of action of the most common antibiotic resistance...presence of an antibiotic resistance gene is a mechanism to culture only bacteria which have retained a...carb and amp, due to their common degradation mechanism. How to choose between the two? Carb is generally... colonies are an issue. Pros: Very stable, interchangeable with Amp, timesaving for transformations. Con...similar relationship as amp and carb (they are interchangeable). Like carb, spectinomycin is the more stable...

...that you will contribute openness, clarity, approachability, and professionalism to team meetings and projects...yourself  standing in front of a crowd and using your charisma to give people hope for some shiny vision on the... you’re more of a leader than you thought! When change happens at your organization, when your boss needs...presents itself even outside of work, consider challenging yourself, take the opportunity, and discover ...to manage these responsibilities. If you are in charge of completing many meticulous protocols or tasks...

...may provide a method for doing just that. While challenges remain in the delivery of these agents, CRISPR...resistance using multiple approaches. Major restaurant chains including Chipotle and Panera Bread have pledged...remain to help recolonize the niche, reducing the chance of an opportunistic infection like for organisms...quinolone resistance, mediated by a single base pair change in DNA gyrase, CRISPR phagemids were cytotoxic .... PubMed Central: PMCID: PMC4317352. 3. Beisel, Chase L., Ahmed A. Gomaa, and Rodolphe Barrangou. “A CRISPR...

...developing novel technologies to analyze molecular mechanisms of genome functions such as epigenetic regulation...epigenetic regulation. To elucidate molecular mechanisms of regulation of genome functions, we need to...will markedly accelerate research of regulation mechanisms of genome functions in the hope of developing... which is very disappointing. I hope I can help change the situation in the future. Addgene: What advice... they may be able to achieve something that may change the rules of the game. In this regard, I believe...

... Protocol Exchange Website: http://www.nature.com/protocolexchange/ Nature Protocol Exchange (NPE) is ...your favorite protocols for future reference. The exchange currently boasts over 2,500 protocols covering...life science labs and offers the service free of charge to scientists by allowing companies to pay publish...

...buy that charming fixer-upper house you have always admired or volunteer to lead a charitable cause.  ... your work. Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life... spouse won’t be at every school performance, chaperone every field trip or cheer at every soccer game...your plan to live a good life may be the most challenging assay you will ever have the opportunity to develop...

...gRNA together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb...the group to focus their efforts on SaCas9 characterization for in vivo studies. One of the pitfalls of...genes, they saw indel formation and phenotypic changes as early as 1 week post-injection. Livers from ...jejuni (CjCas9) is the smallest Cas9 ortholog characterized to date. Kim et al. successfully used CjCas9...transgenes might seem daunting, it’s actually a challenge that the field has overcome with split AAVs. In...

...When you think of Addgene, chances are software development is not one of the things that comes to mind...the flexibility to adapt and easily respond to changing needs of the teams at Addgene. Another advantage...to bake a cake. Instead, you go to a store and purchase flour. However, buying a bag of flour is not the...updated in several years) or they make significant changes that make it incompatible with our software. When...Python 2.7 to Python 3.5. There are significant changes to the language and it took us a total of two years...

...in contrast, engages existing gene regulatory mechanisms to block activity and does not work easily in...into genetic compensation, a previously unknown mechanism of gene regulation (El-Brolosy et al., 2019). ...fluorescently labeled, allowing researchers to observe changes to these tissues under different experimental conditions...time and space, and was also used to precisely characterize developmental defects in the one-eyed pinhead...2016. 219-244. PubMed PMID: 27443928. Harland, Richard M. "A new view of embryo development and regeneration...

...share all of these new organisms continues to be challenging. Organism specific viral vector systems would...thus making progress on the NSF “G to P grand challenge.” Creating a productive, new scientific community...So, think beyond Drosophila melanogaster or Saccharomyces cerevisiae (both fine research organisms) -  ...social bonds that they form including genetic mechanisms that control who a female wants to mate with ... first? They are starting by seeing if they can change long legged lizards into short legged lizards.   ...

...counter-intuitive, but I find office work more mentally challenging than lab work at times.” Sometimes a desk job...experience will be a little different. Some of these changes you may enjoy immediately and some of them may ... just take some getting used to. Just remember: change is good. You can experience a real onboarding...to non-scientists can be a fun and gratifying challenge. You’re also getting the opportunity to learn ...eventually. If you approach that transition dreading the change, the adjustment period will be that much longer...

...and Discovery As part of their quest to find, characterize, and develop new CRISPR systems and technologies...proteins that could cut RNA or DNA via a CRISPR mechanism. The weird black pellet's DNA yielded a new CRISPR...Prime editing — the ability to now make any small changes, including base edits, insertions, and deletions...right integrase for this new approach proved a challenge. The old standby of CRE/flp didn’t work – it turns...be independent of gene repair.) This lowers the chance of the insert being “dropped” in the wrong place...

...Addgene’s YouTube channel.   Figure 4: Global viewership of the Addgene YouTube channel, 2023. Views from...protocols. In 2013, we launched the Addgene YouTube channel to add a strong visual component to our educational...is achieved through interconnected incremental changes. It was only after creating a plasmid repository...learned about the importance of smaller, systemic changes, like using alt-text, captions, and inclusive design...

...Monster probes at Addgene Uses of smFPs 1. Multi-channel connectomic tracers Fluorescent proteins such ...anti-FP antibodies. GFP emits light in only a single channel. In multi-color imaging experiments, the second...making them difficult to use in a multicolor/multichannel imaging set-up. In contrast, smFP tags can be...University Press, New York. pp 719. 3.Rizzo, Mark A., Michael W. Davidson, and David W. Piston. "Fluorescent ...4. Shaner, Nathan C., George H. Patterson, and Michael W. Davidson. "Advances in fluorescent protein technology...

...methods measure different characteristics of the solution. These characteristics give us information on ...temperature parameters. For example, we found that changing the annealing temperature of the PCR from 60° ...al. "Systemic errors in quantitative polymerase chain reaction titration of self-complementary adeno-associated...PMCID: PMC3216358. 4. Lock, Martin, et al. "Characterization of a recombinant adeno-associated virus type...