...saves you from having to create a new fly line every time you want to express YFG in a new tissue type (more... new fly with hedgehog under a new promoter each time a new hypothesis is tested. Controlling multiple...running with fly lines that are already available, sometimes you’ll have to make your own. In my next post,...process of generating your own fly lines. In the meantime, be sure to browse the publication references ...expression systems in Drosophila: A synthesis of time and space, Trends Genet. 20, 384–391 (2004). PubMed...

...meetings, we cut the time down to an hour or less and are able to provide more timely feedback on each other...idea why you are there. Make the most of everyone’s time by considering the level of meeting you need. Project...meetings short, informative, and pleasant. Email: Sometimes, the right meeting is no meeting at all. When ...assigned items to keep the project moving. Use time wisely Meetings are for talking, not reading: Send...

...or if you'll be going through small aliquots at a time, the -20°C is your best option, in aliquots no smaller...experiment you’re planning to run with the antibodies time-sensitive, using precious samples, or being compared...reaction. For instance, sodium azide, a common antimicrobial agent added to buffers, can bind to amine groups... of 0.01% w/v can be used as an alternative antimicrobial.  Conjugates And HRP brings us handily to the...for an experiment is easy, but what about those times you need more than one?   Multiplex assays Multiplex...

...promoter driving multiple combinations of WT, codon optimized, untagged, cleavage sites (thrombin and TEV), ... coding genes, tags, truncations, and codon optimizations similar to those found in the bacterial expression...plasmids Finally a set of wild type and codon optimized, full length and truncated versions of SARS-CoV... scientists without performing the full suite of time consuming, costly, and laborious QC for all several...re glad to be able to do so during this critical time. Visit our COVID-19 Plasmids and Resources Page!...

...are powerful tools to measure real-time cellular reactions to stimuli and as well as changes in cellular...analyte present in the system at a given point in time. For example, if ATP was the molecule of interest... sensors are fluorescence-based and have been optimized for a wide-range of uses to monitor the fundamental...systems, rather than transiently overexpressing at the time of use. Another great viral perk for delivery is...

...the number of genes that can be manipulated at one time. Weissman’s group has made standard protocols available...available and they are working to further optimize those now. They are also developing sgRNAs that will...there is potential there,” he says. "We have to optimize how we can deliver these tools because obviously...mouse, we can’t rely on genetic engineering to optimize humans for CRISPRi and CRISPRa.” Before considering...information about SAM, including a tool for getting optimized guide sequences for activating any human gene,...

...course, real relationships go further and grow over time, but being interested in the same phosphate of your...connections What can you do to get the most from the time and money you are spending on a conference? One ... onsite and eat together because scientists have time to really connect with others whose company they...yourself. Put your phone away! This is a natural time to meet someone since you are both interested in...promised to send a paper, send it. If you had a nice time, send a note to say so. If you read a paper that...

... research and clinical applications. Most of the time, when someone speaks about an affinity reagent, ...large enough that their bulk can be a barrier at times. Smaller, slimmer affinity reagents can have deeper...have them, you’ll need to either re-develop or re-optimize your assay for an entirely new affinity reagent...reagents you use, one thing is clear: it’s an exciting time to be a scientist looking for that perfect fit!  ...References Grumezescu, A. M. (2018). Drug targeting and stimuli sensitive drug delivery systems. Elsevier Science...

...developmental delays or autism spectrum disorder. It is estimated that CTD is the second cause of male intellectual...psoriasis is a chronic autoinflammatory skin disease estimated to affect 0.27 to 4.6 per 10,000 people globally...of the aorta) are two heart defects that are 50 times more prevalent in women with TS. “The goal of our... bicuspid aortic valve at a rate that is 50-100 times higher than women in the general population,” says... the signalosome of different receptors upon stimulation. “To study cellular transduction events, we will...

...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...

...signal at the same time point. E. coli’s BirA* was tested at its optimum 18 hour time point, but one minute...Sometimes it feels like DNA and protein get all the attention.There are numerous ways to detect DNA-protein... interactions in events that occur on a short timescale, such as during drug action or cell signaling....

...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 ...

...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...

... 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 ...

... 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...

...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...

...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...