...industry postdocs have a set time limit. At AstraZeneca, this is three years. This time limit can feel like a...might have less time to explore scientifically since you can’t afford to spend too much time working on something...Pros and cons: The academic vs. industry postdoc  Time: Academic postdocs can be long—5 years or more, ... Also, if you work in industry for any length of time, you’ll know more people in industry, which means...numbers of publications, which may take a longer time to acquire. Learning about drug discovery: Unless...as AstraZeneca, will hire their postdocs for full-time positions should a position open up. Other companies...

...small amount of DNA can be copied many many times in a short time period. PCR was invented in the early 1980s...denaturation, annealing and extension is repeated 25-35 times to exponentially replicate the target DNA of interest...which would slow or stop PCR amplification. Other times, the PCR reaction may not be 100% efficient and ...polymerase chain reaction (qPCR) also known as real time PCR is a PCR technique used for measuring a starting...require a reference or standard curve, which can be time consuming and challenging to get right. ddPCR uses...an extremely useful PCR method that can help save time and effort in the laboratory.  There are two main... different pathogens in one specimen saving both time and effort. Scientists can also utilize multiplex...

... anaerobic conditions. Maturation Time: Maturation time is the time it takes the FP to correctly fold ...Dreiklang). Fluorescent Timers (FT): These proteins change their color over time. Therefore, these can ...can be used as “timers” for cellular processes following their activation. The four main FTs are called ...and DsRed ~10hours. Temperature: FPs maturation times and fluorescent intensity can be affected by the...temperature. For instance, enhanced GFP (EGFP) was optimized for 37°C, and is therefore most suited for mammalian...

...are some tips and tricks you can use to obtain optimum transformation efficiency. It can take 6-8 weeks...pretty important to get this process right the first time. I’ll lay out some guidelines here. A detailed transformation...transform early, your plants will still have plenty of time to produce seeds after you expose the flowers to...you are transforming more than one construct at a time. For detailed protocols, volumes, spin speeds, ...agrobacterium: Getting a transformant takes a long time. Make sure the agrobacterial culture you are growing...cells in your culture will no longer be viable. The time this will take depends on the volume of culture ...inoculated from a colony on a plate will reach an optimal density in 18-24 hours. I then add 0.5 mL of this...

... for structural studies of E3 ligase cereblon Optimized mScarlet-based tags for live imaging in C. elegans.../10.1038/s41467-024-52871-9. PMID 39406745. Optimizing mScarlet tags for live imaging in C. elegans ...for mScarlet3 and mScarlet-I3 that were codon-optimized for C. elegans and included artificial introns...mScarlet3 was 50% brighter than the original worm-optimized wrmScarlet. mScarlet-I3, while initially dimmer... et al., 2024 under CC-BY license.  These optimized fluorescent proteins will be a useful addition...efficiencies of up to about 9% of colonies (102 to 104 times higher than traditional homologous recombination...mechanical forces with PILATeS By Emily P. Bentley Time to get flexible! Alex Dunn’s lab and collaborators...

...loxP sites (For details see Cre-lox blog post). Optimizing FLP FRT through smart engineering Different FLP...first version of FLP discovered has a temperature optimum of 30 °C and is therefore inefficient in mammalian...identification of FLPe which has a temperature optimum of 37 °C (Buchholz, Angrand, and Stewart 1998)....less efficient than Cre recombinase. FLPe codon optimization resulted in FLPO, which shows similar efficiency...the recombination should only occur at a specific time point? To allow temporal control, ligand regulated...and its synapse should be visualized at the same time from two different plasmids this approach becomes...rearranges these synapses in the living brain over time (Villa et al. 2016). Combing FLP and Cre for additional...

... the better (Nat. Biotechnol., 2014). Given the timing, it shouldn’t be a surprise that the CRISPR system...software to make sgRNA design accessible to all. At the time I thought there was room for improvement and a year...iterative design; it is caused by not having the time to assess public response; and, it is the stage ...winner, is there something that we can invest our time into learning and extract the most value? This is...and not general opinion. I could spend a lot of time running through each tool in detail, but that is... genomes of any size and are only limited by the time it takes authors to add a genome to the tool’s repertoire...those who are entering sgRNA design for the first time, it is best to choose a tool based on immediate ...

...one of the earlier epicenters of the outbreak ultimately flattening the curve, it has become evident that... reagents and equipment and the long turnaround times, have led researchers to turn to other technologies... Sensitivity:10-100 copies of RNA per microliter Time: ~40 minutes Equipment required: 37°C heat block... developed so far.  A similar method has been optimized to detect the SARS-CoV-2 virus directly using ...milliliter can be detected – in the zeptomolar range Time: ~1 hour Equipment required: 37°C water bath and...microliter of RNA targets in a 40-minute incubation  Time: ~90 minutes Equipment needed: 37°C heat block, ...CREST) Sensitivity: 10 copies of RNA per microliter Time: Not mentioned in the paper, likely approximately...

... 2022). The oxytocin sensor MTRIAOT enabled real-time observations of oxytocin dynamics in live mice under...here!   Ino D, et al. A fluorescent sensor for real-time measurement of extracellular oxytocin dynamics in...synaptic glutamate response. They generated variants optimized for different applications (v82 has higher affinity...indicator (GEVI) called JEDI-2P (Liu et al. 2022) optimized for two-photon imaging. They use a unique and ...from Gary Yellen’s lab is LiLac, a fluorescence lifetime sensor for lactate developed using a high-throughput... for performance under two-photon fluorescence lifetime imaging. B) LiLac responses are highly resistant...multiparameter screen for accelerated development and optimization of soluble genetically encoded fluorescent biosensors...

...from paper to an electronic lab notebook requires time and knowledge. In a recent article, we translated...your research can be patented? I.e. is automated time stamping (detailed records on who did what and when... when) important to you? Some ELNs record the times and dates of your actions. These records can take ...about every action performed in the lab (user, date, time, action) are required for regulatory purposes in...them.  Conclusion Researchers spend 40% of their time on tedious administrative tasks that drain resources...resources from the actual research process. It does take time and effort to implement an ELN, but on the long ... organized within an ELN already. Investing some time in deploying a software solution that makes it much...

...non-invasive and offers a high degree of control over time and space. Zebrafish have become a popular model...reflex. These traits, coupled with short generation times and high fecundity, make zebrafish ideal for high...neuron more positive or more negative.  Neuronal stimulation: Depolarization When the inside of the neuron.... When startled by acoustic, visual or tactile stimuli, larval zebrafish react by performing a distinct...behavior is not done in response to any specific stimulus. While the escape response neural pathway is well...the overlap between groups.  They found that photostimulating the Kolmer-Agduhr cells was sufficient to ...Kolmer-Agduhr cells on one side of the body required photostimulation in order to drive the full symmetrical response...

...temperatures for long time periods and workflows where CRISPR nuclease activity is desired and time or protocol ...the hot springs My husband, Paul Blainey, at that time a post doc in the Quake Lab, volunteered to do the...joined for half of the trip. Preparations ahead of time included contacting YNP staff to arrange permits... the 3,471 square mile size of YNP, it took some time to map a route that would reach all of the remote...IgnaviCas9 protein is 1,241 amino acids long, has optimal sgRNA spacer length of 22-25 nts, and is thermostable... Cas9 from Parvibaculum lavamentivorans, whose optimal growth temperature is 30°C. Find IgnaviCas9 here...

... changes of these Twitch sensor variants were optimized by testing various linker configurations in a ...screen in rat hippocampal neuron cultures. The optimized Twitch sensors allowed the researchers to see ...he recommends they give it a shot. “Beginners sometimes think it’s complicated, but actually it is very...that the approach he and his colleagues took to optimize sensitivities can now be applied to dozens of ...simple swapping of binding domains, in practice optimizing the sensors involves plenty of trial and error...between different molecules like zinc and calcium. Ultimately, Merkx said he hopes others will find his tools...s like a test.” References Thestrup et al. Optimized ratiometric calcium sensors for functional in ...

...experienced mentor. The lab was very, very quiet. Every time I asked a question, I felt like I was yelling. I...information you will need is about you. It’s worth taking time to honestly evaluate your own style of work and ... help identify your priorities and needs, but sometimes it just takes introspective thought on what’s ... thrive. How big is the group? Has it grown over time? If not, why not? It may not be a bad thing if the...humiliation, accusations regarding lack of effort, intimidating use of discipline or competence procedures Threats..., persistent teasing, name calling, insults, intimidation, sexual harassment Isolation – including preventing...Do lab members work on more than one project at a time or over their stay in the lab? Do all lab members...

...nucleases are promiscuous. In other words, they can sometimes localize to unintended sites with sequence similarity... mutation burden. The CRISPR platform must be optimized to reduce off-targets and patient risk. Hopefully...experiment, at least one will likely be useful. Optimal gRNA selection With some CRISPR experiments, there...can reduce off-targets at a fundamental level. Optimal Cas selection Once you’ve selected a gRNA with ...elsewhere in the genome to reduce off-targets, it is time to consider your Cas enzyme. Even if there are similar... spCas9, has gone through multiple rounds of optimization which have led to more specific variants. HypaCas9...Rabinowitz and Offen). Of note, most of these optimized Cas enzymes prevent cutting at mismatched guide...

...platforms like HaloTag® Fusion Protein, codon-optimized Firefly luciferase genes (e.g., luc2), and NanoLuc...issues as traditional luciferases emit at near optimal wavelengths (Fluc emits at 610 nm) and do not require...’ dim light emissions requires long acquisition times that may be impractical or infeasible for certain... emission peak is at 460 nm thus making it non-optimal for in vivo imaging. The solution is to couple ...quantum yield, brightness and improved maturation time compared to similar orange fluorescent proteins....et al. (5) imaging multiple proteins at the same time in live cells using fluorescent proteins (FPs) can... variants of bright luminescent protein for real-time multicolour bioimaging. Nature Comm. 7, 13718. PubMed...

...working hours (9am-5pm Mon-Fri or overtime and weekends, part time or full time) and what life is like for someone...correctly the first time. These skills are very desirable to employers because they save time and money and ...attracted to science because they enjoy spending time outdoors and with nature. Experience in the field...assessed your skill set and done your research, it’s time to start sending out applications. Check out our...

...where you met and what you discussed. Do this in a timely manner (within a few days or a week) so they can... to build relationships and strengthen them over time.  Things to avoid when following up with your ...connect via LinkedIn with strangers. It’s a waste of time. People that don’t know you, can’t really help you...people who do respond. Don’t call or email multiple times if you don’t get a response. It breaks an important...them and they don’t know me. It’s a waste of our time.  Networking connections can last years This is...ask me about the different arms of this starfish sometime, but as of yesterday, I have 4,700 LinkedIn connections...necessary. Once you’ve talked with a person a few times, if it was a positive experience for you both, they...

...Years later, these classic systems continue to be optimized by Tabor’s lab, making light-controlled gene expression... biological processes. Chemical effectors are suboptimal for a number of reasons, including their potential...high or very low levels. Schmidl et al. chose to optimize these systems to make them more tunable and user-friendly...transcription factors, for which low expression levels are optimal. These tools are compatible with many strains of...et al. also represents a large step forward in optimizing TCSs. Over 75,000 TCSs have been identified in... in bacteria, and with similar optimization methods, many of these systems could be made suitable for ... applications.   References Refactoring and optimization of light-switchable Escherichia coli two-component...

... available amino acids a bit…limiting. It may be time, then, to look towards genetic code expansion (GCE... which non-canonical amino acids (ncAAs, also sometimes called unnatural amino acids) are placed in a ...along with the inherent trickiness of using and optimizing GCE, led to a large toolbox filled with systems...dubious enough that they don’t give it a try. Optimization Why is GCE so complicated? Well, while the techniques...growth temperature) may need to be individually optimized for each of the three factors: the ncAA, the tRNA... synthetases. And, naturally, rarely are the optimization parameters the same for all three components...including their phosphoserine ncAA protocol, to optimize and share, they’re soliciting feedback from scientists...

...combined optical and genetic methods to photo-stimulate mammalian neurons and alter synaptic events: a...rhodopsins and neurons have in common? Neuronal stimulations are triggered by the movement of ions across...viral-infected neurons are subsequently photo-stimulated through fiberoptic cannulas directly implanted.... In particular, they found that optogenetic stimulations of the DA neurons produce behavioral and electroencephalography...could activate appetitive rather than aversive stimuli. In a recent paper, de Jong and colleagues used...neurons of the VTA. Using in vivo optogenetic stimulations, they simultaneously recorded the electrical...PMC3225502. Boyden, Edward S., et al. "Millisecond-timescale, genetically targeted optical control of neural...

...this really sounds like a two-way ANOVA. And this time, it is! Factorial ANOVAs include two-way ANOVAs ...impact of temperature, humidity, sunlight exposure time, and watering volume on total crop production (though...the only things that matter. However, this is oftentimes an oversimplification. If your independent variables...every pairwise comparison within your population? Sometimes this can’t be avoided, but if it can be, you can...example, if your alpha is set at .05, then 5% of the time the statistical test will be incorrect. If you have...high false positives. Control vs. all samples Oftentimes there exists a control group and multiple experimental...working with, graphing data with a histogram is sometimes sufficient to determine normality (or lack thereof...

... a full-time career or a part-time passion, there are many ways to make it work for your time and interests...is science policy?”  I have been asked this many times throughout my short science policy career and this...including fellowships, volunteer positions, and part-time opportunities. But even through all my roles and...academia as tenured professors. There’s never a bad time to start your science policy journey.  In fact, ...Note: the answer to this question may change over time and that’s perfectly normal!  Action Items:  Start...presenting for non-scientist audiences during your time in graduate school through outreach organizations...

...that the optimal buffer for Taq DNA Ligase contains 100 – 200 mM KCl at pH 8.5. This optimization method...condition optimization, will be critical for success. Online tools are available to help optimize ligation... the incidence of annealed mismatched dsDNA. Optimization through high-throughput profiling New England... both the enzyme and the reaction buffer were optimized, resulting in the highest fidelity NAD+-dependent...Reaction Temperature Calculator can be used to estimate an appropriate incubation temperature when using...: 23842410. 19. LeClair, Norbert P., et al. "Optimization of a ligase detection reaction-fluorescent microsphere...

...Before diving into the experimental work, spend some time outlining the construction of the plasmid and all.... It is always a good idea to perform primer optimization, especially if you are having difficulty amplifying... genome, you can recheck your primer design or optimize your primers or PCR conditions. Purifying DNA...increase your chances to have a successful and ultimately faster cloning. Remember to quantify the DNA ... parameters that you can adjust: temperature and time of incubation, and amount of DNA. As a general rule...plasmid:insert), but this parameter has to be optimized according to the strategy adopted, and to the ...and if the strain is growing fine, it is probably time to prepare new plates with a fresh stock of antibiotic...

... do you have to work with difficult people? Each time I ask this question at my workshops, I get nods ...impolite, Complainers, who spend most of their time moaning about their troubles, Silent or unresponsive...supervisors: Hands-off super busy types, who never have time to mentor you, and Micromanagers, who question.... You can save yourself and your supervisor some time if you come to every meeting with one or more proposals...Your supervisor will appreciate that you took the time to think of solutions, and will be more likely to...your action item list and set some approximate timelines. Be sure that you keep your commitments, and if...your point across. Some supervisors might try to intimidate you with negative comments (hostile-aggressives...

...conversation to pass the time in a job that wasn’t going anywhere, but it became a legitimate brain exercise in...still flip to that page in my lab notebook from time to time. I wrote in large letters “Going for it!” We...there very few jobs available. So, we spent a lot of time talking about what we could do to make ourselves...weren’t restricted to science jobs. Keira’s an optimist, I’m a realist, and together we make a pretty ... 2014. Outside of the lab, Keira enjoys spending time with her husband and two Jack Russell Terriers. ...

... and significant lengths of time, the stability of fluorescence over time is crucial. For instance, the... century, it can be overwhelming to select the optimal dye or fluorescent ligand when designing an experiment...intensities, and the best practice to determine optimal DOL is through several small-batch experimental...can be mitigated through careful selection and optimization. Fluorescent experiment types: Different types...super-resolution microscopy techniques, such as Stimulated Emission Depletion (STED), and Stochastic Optical...non-fluorescent states in response to a specific stimuli, which leaves only a small volume of fluorophores...technically cell-permeable, its use in live cells can sometimes result in cellular toxicity due to the permeation...

...on the Addgene Blog Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User) The Challenges of Cell...colonies of antibiotic-resistant cells; during the time required to grease, place and fill a dozen cloning...dilution cloning into 96-well plates, is extremely time- and resource-consumptive, since only wells with...well-spaced clones. For rapidly growing cells we sometimes use TPP peelaway T-75 flasks to provide a greater... in outgrowth. Depending on the cell type, we sometimes add a drop of serum to each well in the 48-well... the University of Maryland Medical School in Baltimore. She is particularly interested in signaling peptide...

...articles.  Here's what you'll find in this post: Optimized prime editing in dicots Self-labeling tags for...Antibodies for neurodegeneration research   Optimized prime editing in dicots By Emily P. Bentley Prime...combining several approaches (Vu et al., 2024). Their optimized strategy uses new combinations of PE protein components...recombined prime editing tools PE2max-NC and ePEmax3 optimized for dicots. NC: nucleocapsid RNA chaperone, RT...Song, Y. J., Nguyen, T. H., & Kim, J.-Y. (2024). Optimized dicot prime editing enables heritable desired ... their propagation along dendrites for the first time (Figure 3). The team also used AAV9 to deliver ASAP5...expresses all the components of the biosensor in optimal proportions from a vector backbone that allows ...