...sets in. Most of the time, that’s enough–but if it’s not, I can always take more time to check my notes!...share what I've learned along the way. Own your time When I was an undergrad, I was assigned a paper ...lesson: Your presentation will be better if you take time when you need it, rather than trying to push through...resources on the Addgene blog Transferable Skills Guide: Time Management Transferable Skills Guide: Conflict Resolution...

...intentional introduction of the Cre recombinase. This is a time-tested system but does require knock-in of the LoxP...genomic site, or randomly integrated (once or many times) into the genome. Make sure to choose an appropriate...Piggybac dCas9 SAM vector! On/off control all the time If your experimental system requires tunable control...turn expression on/off for short or long periods of time. Thus, there is a lot of overlap with the systems...site-specifically knocked in. This can be labor intensive and time consuming. If your native protein doesn’t require...

...monitor their abundance and clonal dynamics over time using next generation sequencing (NGS). The library...implantation. Their goal was to figure out how to optimize protocols for in vivo screening with lentiviral...of each barcoded clone can then be monitored over time by sequencing the barcodes in the population (all...tool in their own work. She says, “I spend a lot of time packing, shipping, and sharing protocols one by ...Many thanks to Carrie and Novartis for taking the time to talk with us and for providing this powerful ...

...to appropriately time the injection about two weeks before the behavior test. This time window between ...measuring locomotion. Mice may need to be given some time to adapt to a testing apparatus. General stress ...cleaned thoroughly between animals as olfactory stimuli that were left behind from a preceding animal (...repeated staggered injections may be considered. Timing: High expression levels of the viral vector trangenes...

... a precursor molecule is made, modified (sometimes many times), and isolated afterwards. To engineer these...other sources; second, characterizing a part or optimizing a TU is much easier when you can quickly test...assembly. These pathways can require a lot of optimization - which is why MoClo is so attractive! It allows...delivering them to these organisms via another optimized platform. Gene Stacking As the name implies, gene...

... for every new prime editing application. And optimizing its sequence is important, as prime editing efficiency... with prime editing, you’ll need to design and optimize a pegRNA for your particular edit. These tips ...with G/C content outside this range can still be optimized (Anzalone et al., 2019). Test different lengths...2019). Figure 2: Starting lengths for optimizing your pegRNA primer binding site (PBS) and reverse...tool-specific best practices. Here are some tips for optimizing your tool of choice. Nicking sgRNAs PE3 and ...

...an enormous amount of wasted money and time, with an estimated $350M/year lost on low quality antibodies...remix the songs, the CDs could get scratched over time, and it was difficult to keep track of growing collections...molecularly defined antibodies that do not change over time, scientists will know exactly what antibody they...lack sequence data, and may genetically drift over time. Antibodies made from animals are limited in supply...

...publication timeline I think we can speak for all the authors on the paper when we say the timeline to develop...complete the project on such a short timescale. Our article ultimately reported the development of a rapid...the American-lineage of the virus. For the first time, we have also demonstrated our technology on active...

...An estimated 320,000 viruses can infect mammals. Even more abundant are the Earth’s estimated 1031 bacteriophages...of the infectivity of your viral stock, save you time, and allow you to standardize your experiments later...References 1. Anthony, Simon J., et al. "A strategy to estimate unknown viral diversity in mammals." MBio 4.5 ...Stability of a recombinant adenoviral vector: optimization of conditions for storage, transport and delivery...

...on the Addgene Blog First time CRISPR user? Check out our tips for 1st time CRISPR users Learn how to ...factors, can increase a gene’s transcription. Sometimes enhancers are located thousands of base pairs ... interactions at one location in the genome at a time (Liu et al., 2017). Recently the Xu lab developed...epigenetic modifications, we can start to build a timeline of events that surround changes in gene expression...

...viewed 1,904,462 times, and the CRISPR 101 eBook has been downloaded over 72,000 times. Our CRISPR blog...2012 to 2023; data are smoothed using locally estimated scatterplot smoothing. (C) Number of requests ...Homology Directed Repair has been viewed 219,289 times, while Validating Your Genome Edit has 76,011 views...Addgene repository have been cited a total of 23,006 times, including 14,677 unique citations.  CRISPR is a...

... Attempts have been made to optimize their fluorescence, which ultimately led to the engineering of the...been achieved with these sensors, but further optimization is necessary for these to be as effective at.... The pro of this system is the kinetic response time from signal input to readable output, which generally...swaps capsid proteins from different viruses to optimize them for targeting your cell type of interest....

... 2016; Duret et al., 2019). But the field has sometimes seen controversy, and the interactions of organic...response to a ~25 mT magnet. Gray regions indicate times when the magnetic field is on. Image reproduced ...response to a ~10 mT magnet. Gray regions indicate times when the magnetic field is on. Image reproduced ... into a variety of optogenetic tools, they’re optimistic that MagLOV can be similarly adaptable into future...Dodson, C. A., Wallace, M. I., Mackenzie, S. R., Timmel, C. R., & Hore, P. J. (2016). Chemical amplification...

...been widely used in biological research for a long time, chromoproteins are just now being developed for...identification is a good place to start before taking on more time consuming and expensive methods. While techniques...manipulations start. Anthony Forster’s lab used codon optimization programs to engineer 14 chromoprotein sequences...lab determined the color intensities, maturation times, and fitness cost of expression. They found that... lab developed as an acceptor for fluorescence lifetime imaging microscopy-based Förster resonance energy...

... enable biosynthetic pathway engineering and optimization. However, current best practices for assemblies... often cycled between 37°C and 16°C to provide optimal reaction temps for both the restriction enzyme ...lacI/lacZ cassette (designed by NEB for use in optimization of assembly systems) to produce a blue colony...improved Golden Gate performance, a series of optimization experiments for these more complex assemblies...    References 1. Vladimir, Potapov, et al. "Optimization of Golden Gate assembly through application ...

...nature. Full disclosure: Zimmer also spends some time on Addgene’s role in the rapid dissemination of ...walk down memory lane for me. A brief window in time “The most obvious future consequence of the undermining...effect it's had on science in a very short period of time. Though climate change certainly still is and will...The State of Science have not changed in “COVID times,” but the pandemic has certainly exacerbated them... of science that I’ve been thinking about a long time. For the scientists who like to learn everything...

...obtained with commercial kits, but in less time. The authors estimate that a gravity-based column plasmid maxiprep...followed by a short centrifugation for a total of four times before a wash with ddH2O and then ethanol. After...yields. Additionally, columns regenerated up to five times had similar DNA yields as fresh columns, suggesting...suggesting that columns can be reused at least five times. DNA purification kits don’t come with enough buffer...

...buffer, it’s time to quantify it. There are several options to choose from depending on the time, reagents...incorporate regular feeds during this incubation time to maintain cell viability and increase protein ...protein stain, such as Coomassie blue. Though more time-consuming, this assay will also measure the purity...stored at 37 °C for a period of two weeks. After this time, quality control assays showed that the majority...

...provide you with more details to help you plan and optimize your own protocols. Here we’ll start with just...freeze your samples in a given state or point in time. You want to halt degradative processes while maintaining...cross-linking that occurs depends on the incubation time and temperature, so you may be able to recover good...protocol that usually works in your system and then optimizing from there is a good option.  Consider your target...examples to help choose which fixative to use!) Ultimately, the only way to know if a method will work is...

...able to expand evenly in all directions to several times its original size. Depending on the protocol, samples...gel recipe used, ambient temperatures, incubation times, and the physical properties of the sample itself... keep distortions to a minimum with protocol optimization and careful execution, but they cannot be entirely... continuous lines in pre-expanded samples can sometimes take on a fragmented appearance post-expansion...Rizzoli SO, Danzl JG (2019) A practical guide to optimization in X10 expansion microscopy. Nat Protoc 14:832...

...and its related fluorescent proteins, which are sometimes used as tags for detection; however, those are...of instability. Prokaryotes can also have a hard time folding eukaryotic proteins. You can get your bacteria... all these tags is that many of them serve a one-time purpose, and you don’t necessarily want them to ...order to use them effectively. One of the best optimized is the tobacco etch virus (TEV) protease. A TEV... overview of why scientists use tags, with a few time-tested tags and fusion proteins as examples. The...

...those engineered DNA-binding molecules for the first time and named the new technology as engineered DNA-binding... applications, for example. However, at the same time, it's fun to be involved in these professional activities...very different cultures. In Basel, I felt that the time flows much slower, like the flow of the Rhine River... incubator for me. In Japan, of course, it is optimized for living for Japanese, which is not trivial,...to publish good publications in a short period of time and may need to do some fashionable projects. However...

...observe the beauty of embryo development in real time. I was already a biology geek, but this formative...developed for each organism, but researchers are optimistic that this can be done for most, if not all, of... to be published, but to engage in ongoing, real-time dissemination of information and materials. Many... branches of the evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (...also return to their nests so can be studied over time. They recognize individual faces (I guess of other...

...Alice Ting have used one biotechnology tool to optimize another. The authors have created a yeast-based...team used other molecular techniques to introduce time modulation of the TEV cleavage and improve utility...2019. https://doi.org/10.1038/s41592-019-0665-7 Optimizing fluorescent tools for labeling structures and...localization of biomolecules within cells. However, sometimes these fluorescent proteins might not localize ...variants of the calcium indicator jGCaMP7, that are optimized for excitation wavelengths above 1,000 nm and ...

...the workload and time commitment for users. This method also requires more optimization and controls when...for a secondary antibody incubation step, saving time. Moreover, since secondary antibodies can cross-...cross-reactivity.   While the method is simple and saves time, it also has several drawbacks. Direct ELISAs generally...antibodies of the same isotype.     Specificity and timing are the major disadvantages of the indirect method...

...working on one process at a time, and testing a small set of data each time a new process was added, we...connected, as this enabled us to rapidly iterate and optimize our pipeline. After reviewing and testing several...path forward was clear: to build one process at a time and connect each subsequent process to the existing... the compute resources when needed, for faster runtime and decreased cost compared to purchasing and maintaining...

...try to keep discussions on-topic and, at least sometimes, civil. r/Science in particular hosts a variety... a description of our AMA and Joanne’s preferred time to start answering questions. r/Science has pretty...alone!) for those going through the really tough times. Getting on a good science career path can be challenging...scientists in training is abundant, helpful, and sometimes excellent. Finally, as I said in the AMA, it’s...Scientists Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments Additional Resources ...

...Church. "Genetically encoded sensors enable real-time observation of metabolite production." Proceedings... as most translation initiation sequences are optimised for a particular system. The Alexandrov lab at...specific alleles in microbial populations through time, Christopher Marx's lab has engineered and validated...quantitatively determine allele frequencies across timepoints or populations. This kit consists of a 48 plasmid...reveals an ordered progression to resistance." Antimicrobial agents and chemotherapy 57.11 (2013): 5373-5383...

...restriction sites (or lack thereof) nor a serious time commitment. Another advantage is that this process...generate wild type and mutant constructs at the same time, rather than sequentially.   How does Gibson assembly...screen a large amount of colonies. Excluding the time it takes to get primers, you can have your construct...their manufacturers tout great efficiency and low time commitments. These kits also come with specific ...such as increased efficiency, shorter incubation times, or the ability to accommodate smaller fragments...

...regular post-holiday routines, why not take a little time to appreciate the humor in science and in the lab... all a place that encourages people to take some time out every once in a while – just read Melina Fan... people in a better mood. Laughter itself can sometimes even count as demanding exercise, and, of course... you that laughter really is a great use of your time even as that lab bench is calling, let me share ...