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...thus individual laser stimulation is essential. In terms of the laser stimulation itself, the output of...control one or two lasers are also available. Optimal stimulation parameters must also be determined empirically...the dorsal/ventral location of your region of stimulation (the z-axis bregma coordinate). Because the light...it enters tissue, placement above the site of stimulation guarantees that the majority of the opsins will...should be about 0.4-0.5mm above the site of your stimulation. The scatter effect is also an important consideration...during the entire protocol, otherwise optical stimulation will be erratic, opsin activation/inactivation...able to purchase one pre-made instead). Laser stimulation         In the video above, Channelrhodopsin...

...have found it saves analysis time to remove this default setting. Timesaving Tip #2: blastx searches are...option to drastically reduce waiting time for search results. Optimizing blastp searches Depending on the ... clones, try these tips to make the most of your time and sequence! You might also enjoy seeing how our...discrepancies. The blastp and blastx programs are optimized differently and you may want to select one (or.... We will delve into these differences below. Optimizing blastn searches On the Standard Nucleotide BLAST...fastest BLAST search to perform and will save you time compared to other types of BLAST searches. If you...may be the most comprehensive for searching. Timesaving Tip #1: If you know the species that your sequencing...

...promoters). Conduct a time course experiment in which you monitor expression at various time points (e.g. every...weeks over 8 weeks) in order to determine the optimal time period between viral injection and commencement...you attempt to stimulate the cell body or an axon terminal also affects the length of time you may need ...like wildfire and for good reason. For the first time in the history of neuroscience research we have ...circuit of interest in promoting reward is a self-stimulation experiment wherein mouse initiated optogenetic...in a reward sensation and subsequent repeated stimulation. A spatial memory task, on the other hand, would...opsin of choice in the brain region you wish to stimulate 2) Fabrication and implantation of the optical...

...Yan labs developed MitoTimer plasmids. MitoTimer proteins are derived from the Timer protein - a mutant...occurs during Timer protein life due to a form of oxidation (dehydrogenation). MitoTimers are composed ...has used an inducible MitoTimer reporter using a Tet-on system (pTRE-Tight-MitoTimer) to show its usefulness...Bomati et al., Sci Rep 2014 Jun 27;4:5469. MitoTimer plasmids to report mitochondrial turnover ...color from green to red when it is oxidized. The Timer tool has already been used by several groups to ...composed of a Timer protein fused to the mitochondrial targeting sequence of cytochrome c oxidase subunit VIII...changes in mitochondrial turnover and transport. pMitoTimer from Yan’s lab, which is constitutively expressed...

...reason we sincerely like spending time together is that when it’s time for fun, everyone is confident that.... No, we’re not all BFFs. Yes, sometimes teams disagree. Yes, sometimes work is a bummer when you’d rather...little neurotic, but hey- sometimes fun needs to be had in an orderly and timely fashion. 9. Ask for help...founders and early employees spent this remaining time by elaborately pranking each other, dressing in ...keeps all of us quite busy. Though there still isn't time for goofing off, the company culture here at Addgene...weekend was the annual Addgene Summer Picnic - a time when Addgenies and their families join together ... it was just beginner’s luck, but I’m feeling optimistic for the next tournament). Minus a little rain...

...usage bias - codon optimization and the expression of alternative tRNAs Codon optimization With low cost DNA... host online codon optimization tools. Keep in mind that, just because you optimize a gene with one of...gene that’s been codon optimized for a particular organism, this will sometimes (but not always) be noted...tRNAs If you don’t have the time or the funds to synthesize a codon optimized version of your gene of interest... Pechmann and Frydman found that tracts of non-optimal codons are associated with specific secondary structures...desired expression host. This is known as “codon optimization.” Though simple in theory, this is not as easy...Luckily, many researchers have created codon optimization algorithms and DNA synthesis companies such ...

...on these findings. Most often we suggest estimating the time it will take to secure your first (R-level...should take the following steps: 1. Define your timeline Start with your thesis in mind. Imagine the kind...R-level) research grant.  Set your timeline from there, then start identifying the resources needed to accomplish...accomplish that goal. 2. Identify the tools in that timeline Avoid writing down prices as you create your list...these tools as possible, this even includes clocks, timers, and phones.  As for reagents and consumables (...”, “weaning”, “colony”, etc. This will help save time by screening out irrelevant candidates, and, more... creates a self-assured discussion when it comes time to decide what resources to keep, and which ones...

...this time and what your needs actually are: Maybe you only need 15 minutes of the two hour time slot,...rest of the time to collect her data. Perhaps you both have an issue with childcare timing pickup. Whoever...available after-hours time slot.  You were hoping that your lab mate would have time to make comments on...successfully!), the process felt unnatural and intimidating. I felt unprepared. Mainly, I thought of negotiation... by people wearing business suits discussing multimillion dollar contracts behind closed doors. ...buffer this week?). Scientists negotiate all the time over everything from which experiment to do next... argument but felt bad afterwards). Or worse, sometimes there’s no agreement at all and everyone leaves...

...with validation and optimization. Although there are different ways to ultimately achieve success, we ...that looks effective. Ultimately, the perfect “high titer” is one that optimizes gene expression where...you’re ready to start selecting, testing, and optimizing three key variables of an AAV vector: serotype... your AAV system and b) steps for testing and optimizing your serotypes, titer, and promoter. Note that... gene expression, but can become methylated over time, resulting in decreased expression (Damdindorj et...). They are useful when you need to control the timing of expression in your experiment. Some inducible... to define high titer, then, is this: It’s the optimal number of viral particles per volume required to...

... wanted to hit out of the park, part time work, bills, (at times) a social life and rest. I need it just...think of a time you’ve lead your team in a defined direction in the past. Maybe there was a time you noticed...   What is a transferable skill? Time management. I needed it when balancing a handful of demanding courses... other not-so-technical skills you’ve built over time are only applicable to those positions you have ...you’re guaranteed to get the job, however, many times the hiring team will also want to examine not just... your potential manager wants to hear a specific time that you thought outside the box to solve a problem...not being tapped at the moment? You can take this time to step up and develop your nascent leadership skills...

...largely replaced by real-time PCR to answer the same experimental questions. Real-time PCR detects and quantifies...are replaced by improved versions that save both time and money. Yet, knowledge of historical techniques...evolution. Southern blotting and quantitative real-time PCR Southern blotting, named for its inventor Edward...requires a large amount of high-quality DNA, real-time PCR has several advantages including easier automation...the amount of DNA used saving the researcher both time and resources (3). Still, researchers generating...blots have a lower false positive rate than real-time PCR (4). Restriction mapping and modern sequencing...to studying DNA sequences helped researchers at a time when sequencing was still in the early days of development...

...single guide RNAs (sgRNAs), are time-consuming, have been typically optimised for immortal/genetically-corrupt... You’ll need to optimize csRNP delivery for each cell type you work with (optimization includes selection...protein. To study 20,000 human proteins, this would be time-consuming, expensive, and not to mention frustrating...RNA and donor DNA design, and implemented our optimised protocols into a medium-throughput pipeline for...now use for IP and ChIP assays. Tag 96 genes at a time using Custom Synthetic cr:tracrRNA/Cas9 Ribonucleoprotein...homology arms (Figure 1). In the first step of optimization, we started off by using custom synthetic two-part...various other suppliers with a typical turnaround time of 5-7 days. The crRNA and tracrRNA are annealed...

...check myself sometimes to make sure I am delegating well, but it does get easier over time! Communicate...management skills can be taught, but it will take time so be patient and keep an open mind. First let’...through doing things. Had I realized that at the time I could likely have retained this talented undergrad...First, it soon became obvious that I didn’t have time to do everything as I planned and I was missing ... be trained, which may seem like a waste of your time at first. After all, you know exactly what and how...faster to do it yourself? Training certainly takes time - so plan accordingly -  but think of it as an investment...investment: once the person is trained you will have more time to focus on what you should be doing : writing a...

...they have time to answer outstanding questions you have, well before experimental crunch time. Don’t: ...needs. Remember, your bench mentor’s training time is time set aside just for you – so do your best to ...progress. You will also probably have a lot more face time with your bench mentor than your scientific mentor... it shows you are motivated – plus it’s a big timesaver for them if you come to a training having already...and/or receive help until you’ve already started a time sensitive experiment. Ready to fly solo? Let your...attempting a technique on your own for the first time. They can then plan accordingly in case you have... In the latter case, you can say “I usually need time to gather my thoughts, but always have questions...

...attend sessions with.” These sentiments were echoed in the other testimonies we gathered. The program definitely...The sheer scale of a large conference can be intimidating. And it can be exacerbated when everyone seems...know one another, but they don’t know you. First time attendees and those attending alone often feel this...While I had been to the General Meeting seven other times I had subsequently changed fields and my cohort ...I didn’t know anything about these people at the time, but I met some very accomplished scientists and... a mentor who had attended the meeting multiple times before and would help the Singletons plan for and... and Singletons can accelerate the networking timetable for the Singletons in their bins.  The next component...

...the threshold at which salaried workers receive overtime payment for working more than 40 hours per week... level (the current average postdoc salary is estimated at around $45,000, but as I’ve discussed elsewhere...hours that these scientists work and paying them overtime accordingly. Read Additional Career Posts on the...2016, Secretary of Labor Thomas Perez updated the overtime regulations of the FLSA. The exemption salary ... to be exempted from the need to pay employees overtime, salaries must be above this level, and certain...Fair Pay for Postdocs: Why We Support New Federal Overtime Rules, made clear that institutional arguments...organizations across academia). Increased postdoc wages, overtime tracking? What are institutions doing? Institutions...

...sending out these questions ahead of time so that lab members have time to prepare their thoughts. Then at...believe that showing something different from time to time breaks up the monologue a little and gives participants...forced to work from home and/or be in the lab at odd times, new strategies can make socially distanced work...creating space for discussion at the end of each talk - time for free interaction and brainstorming. This could...take minutes that can be accessed by anyone at any time. This document can be shared and serve as a springboard... Take brain breaks Meetings can get very long sometimes, and interaction fades away the longer we are ...take breaks. After 1 hour (or another designated time) of meeting, someone could raise their hands or ...

...since hiring opportunities arise at various times. If the timing and interests align for both parties, he...completely happy at work all of the time, the goal is to spend as much time in the "zone" as you can. You know... each other to be physically present at certain times during the day? We asked three lab heads how they...about his or her interests, and spends one-on-one time with the current members to share short-term goals...provide a solid foundation on which to thrive during time spent as a graduate student or postdoc. Although...research field is important, the end goal during this time is to be trained to think and execute tasks like...choosing direction of a project. If you expect more 1:1 time with the advisor, you should choose accordingly....

...an 11-page Cell paper - and sometimes, it might actually take me less time to read the book.  Intuitively...second post in our Summer SciComm series and it’s time for one of my favorite scicomm bandstands: cognitive...up is this: It takes me about the same amount of time to read a 100-page genre fiction novella as it does...Learning, using, and retaining new vocabulary takes time and effort (otherwise it would be a lot quicker ...concepts are easier to grasp than others. But sometimes we’re unaware that we are leaning on an unspoken...connecting information and a logic to the types and timing of questions that are asked. Many things, therefore... and then build up from there. In every project, time, attention, and/or length restraints mean you will...

...reaction, and the timing and number of harvests will all impact the titer. Importantly, optimized conditions ...initial optimization; cell number, media volume, and plasmid concentrations will need to be optimized prior...Additional plasmids will also affect experiment optimization. Before beginning an experiment, it is recommended...additional plasmid for transfection can make these optimization experiments a bit more challenging. With these..., for your experiment. To improve titer, first optimize transfection conditions. As mentioned above, plasmid...efficiency. For some transgenes, transfection optimization simply is not enough to get a sufficient titer...glycol (PEG) precipitation. All of these methods ultimately isolate the viral particles allowing you to concentrate...

...determines the signal’s strength. Exposure time is the amount of time the sample needs to be under light to...compare exposure times of our samples. A prominent, strong signal has a shorter exposure time than the negative...2015), causing a huge waste of precious samples, time, and funds – up to US$350 million in 2015 alone ...molecules), reliable (wouldn’t perform differently over time), compatible with the desired applications (such...strength is based on binding specificity and exposure time. We first have a negative control which does not...goldilocks antibody selection process The exposure time of the primary antibody compared to negative control...to discern the signal. If the exposure time is long (e.g. > 1/7s) then the signal is weak and vice versa...

...discussion? Or maybe the one that goes way over time so there is no time for audience questions and an audience... negatively affect timing and the liveliness of the discussion (see below on timing). Get good resumes...encourage this. Timing is everything Many of my suggestions are meant to keep the event on time. Nobody likes...on a stage if the room is very big, but for an intimate event, put the panelists on the same level as ... show. The moderator must be very active at all times keep things moving and to prevent any one panelist... to introduce themselves and, voila, 1/2 of your time is gone with no interesting discussion. The audience... part of the publicity. Good panels do not waste time on long introductions. The moderator asks questions...

...maintain fidelity with the stimulus. For example, if you want to stimulate at a fast frequency without...very different responses to long vs. short stimulation times or frequencies (Zhang & Cui, 2015). You’ll...that induce conformational changes with light stimulation to either activate or inhibit a specific receptor...the cell to be excited over a sustained period of time (Gong et al., 2020). Light-gated ion channels are...on and off rapidly with light to control precise timing of the perturbation. Other tools are designed to...cryptochrome 2 (CRY2) (Zhang & Cui, 2015). When stimulated with light, the endogenous proteins activate ...experiments.  Wavelengths Most vectors have an optimal wavelength to use but can be activated by a range...

...Harvesting timepoints will depend on your specific virus, so this is something you may have to optimize for ...increasingly accessible and are useful for saving time and avoiding potentially costly set-ups. However...in-house. High-titer adenoviruses are difficult and time consuming to produce, so most labs opt to purchase... kb, while lentivirus has a capacity of 8–10 kb. Time Viral production in mammalian cells is relatively...taking your first round of virus, called P1 (or sometimes P0), infecting new Sf9 cells, and getting an amplified...eliminate dead cells and cellular debris. While adding time (about three to four days), this second round provides...like lentivirus. You can even produce a P3 virus! Time Sf9 cells will start producing virus within a day...

... editing for a few months. More time is required for its optimization and development before it can truly... "Final Verdict", is that more time is required for NgAgo optimization before we can make any major conclusions...PAM sequence might not always be available at an optimal distance from the target site. For example, NGG...been requested by labs around the globe nearly 400 times. Researchers have been excitedly testing NgAgo's...method and 65% wanted to wait until someone else optimizes an NgAgo protocol for their favorite model system...respondents suggesting that NgAgo can indeed be optimized for genome editing in mammalian cells. However...indicates that it is very tricky to use and fussy to optimize. One reason could be the instability of the supposedly...

...interview was intimidating to me, as I had a hard time thinking about how I could fill this time. I was relieved...spent a lot of time shadowing members of my team to learn what I was to do for my time at Addgene. This...make up the hours I had missed at another time. This made time for me to attend to other commitments, without...The steps of the training process gave me enough time to become familiar with the different types of issues...something called “core hours”. Core hours consist of the time between 10 AM and 3 PM, when most meetings are scheduled...the three hours (outside of the core hours) at a time that works for you. For example, if I needed to ...missing work. As long as my work was finished on time, there were no issues. Another example of helpful...

...purpose and finite goals. For example, when it came time to update how we make and distribute kits, I helped... difficult assay with her eyes closed. How many times have you ever seen a scientific article attributed...meetings and we’ve volunteered our ideas, skills, and time to the cause ever since. Unlike the previous two...The benefits of a team So why is it helpful, or sometimes even necessary, to build a team in the first place... when Thomas Jefferson dined alone.” There was a time in human history when an intelligent, motivated ...diverse group of people to provide that base. Sometimes there’s just too much work If you are one of those...other people who have a stake in the outcome will ultimately improve the quality of the work as well. Social...

... Most of the time, updating these libraries is straight forward. However, there are times where a library...genders, educational, and professional backgrounds. Sometimes this can lead to wildly different ideas of how...blind spots. Blind spots often result in less than optimal user experience and the overall soundness of the...stories, and work on stories during short periods of time called sprints. The team is mindful of the number... decide how to best approach it in our code.  Sometimes while thinking through these steps, we discover...code is written, we are able to use our developers’ time efficiently. While writing the code, we may need...developing new features and fixing bugs: we take time during each sprint to do some maintenance work on...

...lower than optimal drug concentration. Drug resistance is not a black or white issue. Sometimes bacteria...killed by a higher concentration, or sometimes more exposure time is needed for the drug to work… if the...superbug’. Superbugs are often very difficult and very time consuming to treat. Learn How Antibiotic Resistance...naturally resistant but has become resistant somehow overtime. One fantastic example of intrinsic resistance...bacterial populations to antibiotics at unnecessary times, putting pressure on resistant bacteria to over-...When you go to the doctor, it will very likely take time to determine if you have an antibiotic resistant...antibiotic resistance is very likely to develop over time. It isn’t feasible to completely eliminate the development...

...figuring out when to schedule times to write.  Pomodoro technique timer  One time management technique that...made your tables and figures. Now it's time to write. Sometimes, it's hard to get those words out and ...kitchen timer that the technique’s developer used as a university student.   The Pomodoro timer is ...borrow this book from your library or don’t have time to read the book, this list summarizes the key points...schedule, even if you only write for a short amount of time each week, and there’s advice on how to actually...great for generating citations for a few papers at a time. Mendeley Mendeley works much like other popular... Hello PhD Podcast episode 15 "Simple tricks for time management: the pomodoro technique": This podcast...