... the hall, it is always a good idea to take some time to confirm that you are working with the correct...of all the plasmids we distribute. This method is time-intensive, so we recommend a variety of ways to ...enzyme. Consult the manufacturer’s manual for the optimal working conditions for each enzyme. Watch out for...The dyes also run at predicted sizes so you can estimate how far down the gel your bands have traveled ...

...enhanced signal of eNL allows their use for the first time in single molecule imaging. Nano-lanterns and your... al. “Expanded palette of Nano-lanterns for real-time multicolor luminescence imaging.” Proc Natl Acad... variants of bright luminescent protein for real-time multicolour bioimaging.” Nat Commun. 7 (2016): 13718...Fluorescent Proteins Check Out Fluorescent Protein Timers Use Rosella to Study Autophagy Resources at Addgene.org...

... of these “variants of unknown significance” (sometimes called “VUS”) function is unknown. How can we ...acid change based on maps of protein structure. Sometimes this provides a great deal of clarity. But, often... other cellular functions and phenotypes. Since time is of the essence in clinical settings, looking ...that could enable future clinicians to improve real time decisions when rare variants are discovered in clinical...

...world of precision genome editing. When it comes time to (hopefully) validate successful mutation of your... we found these assays frustrating to work with, time consuming, and minimally informative. In this blog...Instead of tracking the reaction progress as in real time PCR, each reaction partition is individually assessed... duplexed primer probe assays often used in real time PCR. These assays can be designed to detect either...

...synthetic biology is still in its infancy. A long timeline from transformation to testing and a lack of well-characterized...plants are being investigated on limited funding and time. While institutions that focus on creative problem...like a lot of specialized science, these tools sometimes have limited distribution. We hope that will change... 2014. Outside of the lab, Keira enjoys spending time with her husband and two Jack Russell Terriers. ...

...goals. These programs also serve as a great first-time mentoring experience. They make mentoring less mysterious... all group members to lead and learn at the same time Broadens the network of the participants Can be ...However, they find it awkward to ask someone for the time commitment of a formal mentoring relationship or...organizations as it brings a diversity of perspective. Sometimes it is helpful to find peers that share a similar...

...some of their time. There are many professional coaches who charge for their advice and time. Consider this... or who your new contact will connect you to. Sometimes, it is not who you know but who knows about you...share their wisdom and they might just not have the time. Thank them anyway and move on. If they say yes,...

...cells (iPSCs) has been around for 10 years. In that time, scientists have used almost all available approaches...relatively simple to implement and doesn’t require time-consuming production of virus. In theory this is...efficiencies and they tend to express for longer periods of time than traditional plasmids due to lower activation...therapeutic applications, but it can take a long time to reprogram cells. In Kim et al, cells were exposed...rigorously assesses pluripotency, but they also take less time to complete. Morphology is a simple and fast to ...

... It’s time for another edition of “What’s New in CRISPR” - where we highlight a few of the newest CRISPR...recently shared some exciting plasmid tools to optimize prime editing systems. Engineered pegRNAs improve...upon knowledge of the biology of the system to optimize its outcomes. In their most recent work, the Liu...and improved editing efficiency. The lab also optimized the architecture and amino acid sequence of the...

...allowed scientists to track multiple genomic loci over time using all the colors of the CRISPRainbow. ...telomere dynamics throughout the cell cycle. To optimize the signal to noise ratio of the system, they ...monitor the spatial relationship of the two genes over time, at a range of distances from 2-75 Mbp. Labeling...Imaging of Genomic Loci in Living Human Cells by an Optimized CRISPR/Cas System.” Cell 155 (2013): 1379-1491...

...Benjamin Schwessinger, a postdoc at Berkeley at the time and an eLife ambassador, had previous experience... themselves were assessed and could improve over time. They achieved this by administering pre- and post-workshop...international team assembled the right people at the right time. Be inclusive and seek solutions by working with... industry partners, not the other way around. Sometimes, due to misconceptions and stereotypes, solving...

... tissue culture dish but can also happen in real-time inside a mouse’s brain.  But the brain is a tricky...localization, kinetics, and spectral properties, must be optimized simultaneously in order to generate a channelrhodopsin...reach the VTA. Mice were then allowed to “self-stimulate” their VTA dopaminergic neurons by triggering ...H134R) expressing mice did not have this self-stimulation response. This result showed that ChRger2 stronger...

...of drawbacks, especially the increasingly growing time from submission to publication. It takes many months...peers in the field, and updated by the authors over time, providing a public record of changes, critiques...communication. Researchers often post a preprint at the same time as, or slightly before, submitting the manuscript...your results, share them as best you can! At the time of writing, Mike Lacy was also employed by the American...

...the easiest cloning methods in terms of hands-on time, as digestion and ligation can be done in one 30...Gate cloning into a modular cloning strategy. Sometimes referred to as MoClo, this strategy uses the Type...efficiently assemble up to six DNA fragments at a time. As with all Golden Gate-based methods, this system...Sequential amplification of cloned DNA as tandem multimers using class-IIS restriction enzymes. Lee JH, Skowron...

... reaching consequences, from wasting time and money on optimizing experiments, to false conclusions.  ...errors prevents future experimental issues, saving time and money for scientists. What is a QC issue? At...sequence discrepancies. While Addgene’s QC process adds time to the deposit process, we believe this to be non-negotiable...

...will update information on the plasmid’s page. Sometimes we’re missing a piece of the plasmid There are... in one complete, circular assembly. Most of the time, these plasmids are returned as one or two partial...the correct sample. The PCR is then performed, optimized as needed, and cleaned up using a standard PCR...make available as part of our OpenBio Repository sometime soon. Stay posted!     References Guerin K, ...

...devastating. At the same time, she has an entirely new path to discover and gives her time to give back to her...as a biochemistry major and a Spanish minor, it’s time for me to start considering my future path. Talking... months, she left school when she was hired full time as a Lab Technician. “I was so tired of being in...

...see growth or decline of interest in a field over time. Creating an alert for a search term helps automate...getting a quick summary of papers before investing the time to read them. Organizations: Accounts for nonprofits...used to filter your search to a specific window of time for publication. To help you quickly identify papers...day, week, or month. A measure of the number of times a paper is shared online, or its altmetric score...

...pairs with improved dynamic range for fluorescence lifetime measurements." PloS one 10.8 (2015): e0134436....orientation of the donor and acceptor proteins is not optimal, FRET may not occur or be detected even if the ...experimental setup as a potential source of error in suboptimal or undetectable FRET, well-characterized FRET...intracellular calcium and published in 1997. Since that time, numerous advances in probe design, fluorescent ...

...into the brain over time (Deshpande et al., 2013). The study found that at early time points, most of the... TVA-expressing cells (Osakada et al., 2013). Ultimately, this allows RABV to selectively infect a genetically... came from nearby neurons. It wasn’t until later time points that longer-range connections were seen. ...

...advice I can suggest, is to give yourself plenty of time when writing a fellowship proposal. Yes, yes, I ...likely been through the process themselves many times and will have useful advice about what reviewers...looking for. Remember though, you need to leave them time to read it, give it back to you, and for you to ...these findings will be worth knowing? This can sometimes be tricky, as most of our research builds on ideas...

...progress in clinical therapy is more fitting, so it is time to update our post. From Mice to Men When this ...cells can be delivered immediately, saving precious time. CRISPR can aid in editing these cells to avoid ... financially), and others we just don’t have the time to talk about. Other treatments currently in clinical...Development Resources on the Addgene blog CRISPR Antimicrobials Treating Muscular Dystrophy with CRISPR CRISPR...

...Research (FoR) symposia held across the USA during this time have revealed that young scientists are aware of...either be paid more than $47,476 or be eligible for overtime pay. For many postdocs this will lead to a raise...relationship, along with, of course, union dues. Time will tell the extent of the implications of this...at institutions, will be challenging. At the same time, there is much opportunity to make headway on long-term...

...crude cell lysate (Benyamini et al., 2023) to save time and cost.  Purification To purify your rAAV from...dose (TCID50) (Zen et al., 2004).  To choose the optimal titering assay, you’ll need to understand how different...non-genome-containing (empty) capsids, thus leading to an overestimate of transducible units in the virus preparation...Ehrhardt, A., & Baiker, A. (2012). Universal real-time PCR for the detection and quantification of adeno-associated...

...experiments, thus saving you time, sweat, and money on extensive optimization efforts. This holds true for...score ranking (Kim et al., 2019). PRIDICT saves time and resources by selecting the most promising pegRNA...Koeppel et al., 2023), their predictive power is sometimes limited across different targets due to variations...

... 200x coverage, meaning that you need 30 to 200 times as many colonies as individual plasmids in your ...colonies, you’ll likely rely on serial dilutions to estimate transformation efficiency (Figure 2).    ... Figure 2: Example of a serial dilution to estimate transformation efficiency. Figure used with permission...temperature (e.g., 30 °C), and limiting your growth time (e.g., 16-18 hours). Finding reference protocols...

...library and characterized them by ELISA, homogeneous time resolved fluorescence, and suspension bead assay...degradation, this protein compartment was used to optimize D-glucaric acid biosynthesis. This MPyV system...way to use the accumulation of indels to measure time. These plasmids with different promoters driving...constructs. zCas9i is a high efficiency intron-optimized SpCas9-coding gene. SlugCas9 is a compact Cas9...

...cloner, you’re likely not intimidated by having to pick the right enzymes, optimize reagent concentration,... cloning is another good option to consider when time is of the essence. This method requires a vector...two to generate templates for the cloning process. Time spent actually doing the experiments are also generally...

...nothing is covering it. Turning on the UV light 3-4 times a week for up to half an hour can help keep your...stop growing (contact inhibition) and it takes them time to recover after they are passaged. This could change...borrowing’ cells that have already been passaged many times and could be contaminated.Learn how Addgene uses... and morphology at high passage numbers (>30-40 times), depending on the cell line so old cells should...

... to them about my interests and my ideas all the time. Once, I had this crazy idea that I needed help ...and departments Graduate school was an excellent time to try new things, develop new skills, and meet ...from the experts in science communication in real time! I follow great online science communicators like... why I still love science. Keep building on your time in graduate school I graduated in the fall of 2015...