...the TSS but you can worry less about sequence optimality because you generally have fewer sequences to...so many possibilities, picking a gRNA with an optimized sequence is of primary importance (more on this...subsequent screening for successful edits. This is a very time consuming process and should not be undertaken lightly... for this technology, and the user may need to optimize numerous parameters to generate the desired edit...have a dozen or so gRNAs to choose from in the optimal location. It is also important to have good information...approximately equal importance in design – an optimized sequence will do little if it is in the wrong ...there are fewer gRNA to choose from, and thus an optimal sequence may not be available. Predicting gRNA ...

...irreproducible results to future work cost both time and money with estimates reaching in the billions of dollars ...apply these findings in their own work ultimately wasting both time and money. The obvious solution to ...lab and the entire research enterprise time and funds. It’s time to start putting them into practice!   ...program beneath it. You can jump to the different time stamps in the description to watch any sections ... body works, generating new therapeutics, and sometimes formulating new biological concepts. However, ...the final manuscript regardless of whether the ultimate results were positive or negative. I can feel ...of pre-registration doesn’t come to fruition any time soon, the Center for Open Science does host a platform...

...At the time when we spoke, Susan was helping cover social media while the Hutch hired a full-time social...directions is sometimes at odds with the space needed to be creative. “You need a lot of time to let you ...you mind process things. Sometimes finding that uninterrupted time to be creative is challenging,” says ...written.” Ultimately, Susan feels she was lucky to have been in the right place at the right time when she...research for a general audience. She then got a full time position with a Washington D.C.-based communications...

...Before you begin In the best case, you know ahead of time that you want to probe your blot with multiple antibodies...sample. Pro tip! Because some protein is lost each time you strip a blot, you should not compare the strength...stripping means more sample loss. If you are still optimizing your system, start with a mild stripping solution...5–30 minutes at room temperature. The incubation time will depend on your blot — antibodies that bind ...! βME is a reducing agent and gets oxidized over time, so it should always be added fresh to solutions...washing is critical. Assess the stripping Now it’s time to check to make sure the antibodies from your first...re-incubating with your secondary antibody. This is the one time in western blotting that you’re looking for a “blank...

...residue peptide to the C-terminus of human codon-optimized  cryptochrome 2 (CRY2) from Arabidopsis thaliana..., a researcher can cause them to interact by stimulation with blue light.  Similarly, light can also be... and is reversible upon termination of light stimulation. CRY2clust enables faster assembly and disassembly...integrated into a Ca2+ -modulating optogenetic system (OptoSTIM1), caused two-fold faster changes in intracellular...Listen to the soCoChR podcast segment Optogenetic stimulation is a powerful tool used by neuroscientists to...artifactual PSCs commonly observed with direct stimulation are largely absent in the soCoChR-expressing ... concentrations in several beverages could be estimated by measuring growth of the ΔguaB pDCAF3 strain...

...was a machine that could make your life a hundred times easier. We’ve certainly felt this way at Addgene...month!), manual transformations were taking a lot of time. Enter the Biomek FXP! We wanted a machine that ... the oppotunity to use the FXP to automate and optimize more protocols in the future - something that ...doing about 50 transformations in the same amount of time by hand! Setting up a basic protocol on the Biomek...purify DNA from many bacterial samples at the same time as opposed to preparing each sample individually...developed a protocol to run up to 192 samples at a time. It follows all the same steps and uses the similar...assistant, but with increased plasmid demand it was too time consuming to complete this process by hand and we...

...-- as of June 4, 2020, COVID-19 has caused an estimated total of 380,000 deaths worldwide (statistics ... of plasmids ordered for each paper. Rough estimation of COVID-19 effect on plasmid requests Before...plasmid request activity around March. A rough estimate shows that activity level in March was around ... of the world encountered COVID-19 at different times. Most notably, China was affected by COVID-19 earlier...lockdown. To allow our model to give uncertainty estimates for each parameter, we added log-normal noise ...Each of these regions was affected at a different time by COVID-19, so we defined the COVID-19 event windows...Pytorch, which gave us the maximum likelihood estimates of each parameter. We approximated the data likelihood...

... studies enabled by fluorescent proteins The optimization of GFP and its derivatives by the late Roger...Roger Tsien changed everything. For the first time, fluorescent proteins gave researchers the ability to ...visualize how cells interact after a particular stimulus. Using an epifluorescence microscope and GFP fusion...example, by harvesting lymph nodes at different times after antigen challenge, scientists were able to...to give a clear view of cellular movement in real time. Two-photon microscopy was the next revolution in...the ability to monitor cellular movements in real time. This technique was first Introduced to immunology...migration and movement by marking cells at a specific time. Photoactivable fluorescent proteins, for example...

...North America. I hope that these perspectives, the optimism they encapsulate, and the solutions they promote...promote help motivate you to spend a little time advocating for science and getting yourself out there for...has been here before.  These attacks on science ultimately galvanized scientists to rally in Ottawa to protest...Institute. He also studies energy markets in his spare time. You can follow him on twitter at @thompn4. New ... Saturday in the tourist-trap nightmare that is Times Square. But I made the sacrifice in April because... entire scientific process.  I've spent a lot of time debating how to fix this.  As scientists, we work...lab where the groundbreaking research happens, sometimes forgetting how to engage with friends, family,...

...growth along the way. Take time to celebrate the lab’s achievements and enjoy time with friends and family...accomplishments? Any tips on making time to relax and/or enjoy time away from the bench? We would love ...their team's hard work? Do succesful scientists have time for fun and life outside of the lab? This final ...”. He also cooks and cleans with his family, “sometimes listening to audiobooks.” Church and his family...family. And as the school year winds down, take some time to have fun outside the lab! How does your lab celebrate...George Church (Harvard Medical School) for taking the time to speak with us throughout this series about how...

...Sciences. In the Yang lab, we’re continually optimizing the use of CRISPR editing systems in bacteria...strains, but also in E. coli B and W strains. The optimized plasmid curing method was much simpler and took...Tatumella citrea, with a total of 70 sites edited. We optimized the strategy for plasmid curing, providing users...ways: In our initial paper, we used MUCICAT to optimize the gene dosage of glucose dehydrogenase and generated...achieve glucose dehydrogenase expression that is 2.6 times higher when compared to the industrial strain. In...obtained a stable and high-yielding strain which had 6 times the yield of the industrial strain. Not only is ...with different gene dosages to facilitate swift optimization of strains. Multiple induction systems to choose...

...retreat to the woods for months at a time, so take advantage of whatever time you have. If you’re like me, you... I use the software to add the citations in real time as I write. Things are going to get a little crazy...it’s practical because there simply isn’t enough time to read every single paper. I also learned to appreciate...Like I mentioned, I also added citations in real time as I wrote, so each statement was referenced even...: initially I felt that I needed a huge chunk of time to sit down and get anything done, so unless I had... I realized how elusive a free two hour block of time was. Eventually, I learned that I was most productive... dozens of drafts of the manuscript. This is the time to transform the document into something cohesive...

...companies), and as far as I remember, it was the first time that I was absolutely convinced it was the company...specialist for 4 years. Read on for insights from my time in technical support including tips, advantages,...required for a role in technical support? It takes some time (several years) to build up comprehensive lab experience...companies have dedicated technical support teams, sometimes comprised of several layers of support: A front-line...incoming requests and answers questions that are less time consuming and/or less technically challenging. Alongside...listen to the many individual stories and get (sometimes refreshing) insights into how other scientists...shortage of amusing reports where someone tries to “optimize” a particular standard lab protocol. For example...

...The average latency period (time from first asbestos exposure to the time of diagnosis) is about 35 years... small, hook-like shape of asbestos fibers. Over time these embedded asbestos fibers cause inflammation...which can cause mesothelial cells to mutate and ultimately form cancerous tumors.  This process is not quick...decades from the first exposure to asbestos to the time when someone starts seeing – or feeling – the first...years, but it usually ranges from 20-50 years and sometimes longer. There is no known safe level of exposure...mesothelioma is often misdiagnosed, meaning that by the time it has caught it has usually started to spread (...asbestos for years, or they are exposed even one time. Doctors and researchers don’t entirely understand...

...Erik L. Snapp and Lindsey M. Costantini. "You underestimate the power of the Dark Side." --Darth Vader in... Fusions Observing a bright signal for the first time after cloning an FP fusion with your favorite protein...fluorescent protein fusions in live cells in real time. Previously, we have described a number of practical...protein function and this could lead to under and overestimation of fusion protein activity. We encourage investigators...starting by using the newly published oxidative optimized palette of FPs (moxFPs) [5]. The moxFPs overcome...characteristics of the moxFPs are comparable to the non-optimized parental proteins. Our recent publication illustrated...fluorescent signal achieved when utilizing an optimized moxFP as opposed to standard FPs. The increase...

... and optimization of the transfection reagents and process. It is important to note that optimal experimental...experiments, the biological controls are more time consuming and ultimately more important. Ideally, each biological...identify opportunities for troubleshooting or optimization within your experiment. Read on to learn more...the independent variable should be consistent over time and from different preparations of the purified ...and were incubated on the same plate at the same time. Although the replicate control is important, it... whole experiment from start to finish multiple times and on different days. Using our example above, ...controls, performing multiple replicates, and ultimately getting interpretable results: Figure 4: Expression...

...The first time I heard about FRET during a journal club, my guitarist brain automatically thought about... pAquaN1 Cyan Mammalian Expresses mammalian optimized Aquamarine mCerulean N1 Cyan Mammalian Express...pCEP4CyPet-MAMM Cyan Mammalian Expresses mammalian optimized CyPet pCyPet-His Cyan Bacterial Expresses CyPet...pCEP4YPet-MAMM Yellow Mammalian Expresses mammalian optimized YPet pYPet-His Yellow Bacterial Expresses YPet...acceptor photobleaching method and the fluorescence lifetime imaging microscopy (FLIM) method. Acceptor photobleaching...not to destroy the donor molecule. Fluorescence lifetime imaging microscopy method FLIM has been developed...measuring FRET. FLIM measures the fluorescence decay time of the donor. When FRET occurs between the pairs...

...where to start. Thankfully, my managers set aside time for me to complete an online social media certification... meetings didn’t seem much more than numbers. As time passed, and I participated in the discussions, I...first official professional presentation.  My first time in the lab! I got to assist production on a protocol...slide design, discussion items, big takeaways, and timing. The sense of trust and comfort I had with my teammates...only a Slack message away.  When it finally came time for me to present, I thought I would be extremely...days I was up to my neck in to-dos, I also found time to go out to sushi restaurants for lunch with coworkers...the dog.  Cuddles at Olive’s office hours! As my time here comes to an end, my worries about making connections...

...complementary C. Optimized base editing with eOPTI-CGBE and cOPTI-CGBE Many optimizations had already been...Surprisingly, the team noticed that this editor sometimes introduced C → G edits at certain positions within...were making similar observations around the same time. (The papers were actually published in the same.... Created with BioRender.com. Beyond UNG: Optimizing C → G editors Now that several labs had shown...multi-step cellular repair pathway that is known for sometimes causing double-strand breaks (DSBs). The Fei-Long...common intermediate of CGBE activity, and they are sometimes processed by error-prone non-homologous end joining...editing, we can expect several generations of optimization before the field begins to converge on a set...

...heterodimer in response to blue light, which will ultimately co-localize the VP64 transactivator with a dCas9... innovative ways. The principle of protein multimerization is seen in many important biological contexts...copies of a protein to a target polypeptide. Multimerization in the SunTag system occurs via antibody-peptide...single-molecule reporter of intracellular processes. Multimerization via the SunTag system also shows strong upregulation...specified cellular processes or be observed at a later time via DNA sequencing or other readout. Cellular memory...large changes in binding affinity with light stimulation. Indeed oLID showed only a two-fold change in...50 fold change in binding affinity with light stimulation2. The Kuhlman lab created two improved Light ...