...antibodies through their HLDA workshops. Let’s use TIM-1 (Single pass type-1 membrane protein), which has... Triton X-100; and methanol, we found that the optimal perm/fix solution depended on the specific antibody...wild-type and the KO population (Figure 1), further optimization can be performed to both maximize the separation...concentration (this can also save money!); and optimizing fixation/permeabilization reagents for intracellular...intracellular targets. Since knockout experiments can be time-consuming and expensive, it may be helpful to find...caused by siRNA is also transient, and as such, the optimal knockdown of the protein target will depend on ... is not efficient at the protein level at the timepoint studied Off-target activities of the RNAi Correctly...

...Addgene would also like to thank Viviana Gradinaru and Tim Shay for their support and input. Resources and ... Hudry et al, 2023; Hinderer et al, 2018). The optimal dose for each particular experiment is determined...systemic AAVs are a good fit for your experiment, it’s time to start selecting and testing systemic capsids!...

...receive different types of stimulation may help to refine and optimize your stimulation parameters as well. ...prefer to spend most of its time in the chamber in which it receives the stimulation (conversely, if the circuit... and stimulate them for 30min to an hour and observe what happens. You never know how stimulation of a...personally interested in include: intracranial self-stimulation (ICSS) and place preference. The lab I work in... electrode and non-specifically electrically stimulating an ill-defined region of the brain. With the ...particular hole (the active hole) induces laser stimulation of the chosen brain circuit, while a nose-poke...different patterns and the animal only receives stimulation in one of the two chambers (see image below)....

...quickly Wet transfer Increased time Handcast gel Increased time; have to make as-needed Increase...costs; increased optimization Normalizing to total protein loading Increased time; increased reagents...from lot-to-lot, and have to be re-validated each time a new lot is purchased. Monoclonal antibodies, on...available for your target! Incubation temperature and time Incubating at a lower temperature reduces background...to reduce background while keeping the assay run time to two days.  Indirect or direct detection method...as needed. However, more antibodies mean more optimization and likely higher background noise as well, ... Good for higher weight proteins Long running time   It’s important to note that Bis and Tris gels...

...Reddit AMA Managing time and energy between work and the rest of life is sometimes more of a struggle ... make a change. #2 Thou shalt sometimes put work ahead of life Sometimes, work has to come first. That... for you, but most days go home at the same time. Use time management tools and tactics to plan your work...let my daughter skip school a couple of times. Be sure to add time with your family, for yourself and for...energy and attention. The goal is to maximise the time you are enjoying both work and personal life. There...struggle and sometimes less. When we are lucky, we are “in the zone” and everything seems to fall into place....commandments for making it all work – most of the time. #1 Thou shalt find enjoyment in your work If you...

...reaction optimized. You’ll want to run gradients for optimal annealing temperature and optimal primer concentration...chemistries available and they can be the more optimal choices at times. For example, chemistries that rely on.... The Ultimate qPCR Experiment: Producing Publication Quality, Reproducible Data the First Time, Trends...more in-depth tips on probe chemistries, primer optimization, controls, and analysis.  As a rule of thumb...copy of a transcript. But this sensitivity can sometimes cause as many issues as it solves, so understanding... should be between 100 and 400 nm and must be optimized experimentally.  Probe should have a melting temperature...appropriate reference genes - and it is well worth your time to do one (Biassoni, 2014). At the bench As a highly...

...asked that question today, because I sometimes wish for a stretch of time that doesn’t involve critical thinking...more mentally challenging than lab work at times.” Sometimes a desk job is defined as sitting down at a...how much of that time you’re actually spending doing work. Yes, you may legitimately need to be in the...because of the nature of your time course, but you’ll inevitably have downtime and plenty of it. That’s not...can volunteer their time to share their knowledge with the community (and sometimes conduct their own independent... being the first, and for a very brief period of time, the only, person to know a brand new fact is often...recently who referred to her lab as her family three times during a 10 minute conversation. Leaving a tight-knit...

...block of time and a quiet space where you can address the issue head on. Prepare for this time as if it...need ahead of time and mute your notifications during. (Boyes, 2019) If you’ve taken the time to do this ..., set a timer for five minutes and come up with as many solutions as you can in that timeframe. Tell yourself...you can, set aside time to prepare for future challenges. This doesn’t have to be time-consuming or even...problem-solving process?  You may want to start by carving out time to tackle it. It’s easy to get distracted by an ...hire more people? What would you do with unlimited time? Another strategy for your brainstorm session is...37% higher in originality. It may not be easy to optimize virus production with roller skates—though I could...

...printed your poster and practiced a number of times, it’s time to expertly show off your science.  Prepare...of thought into designing your poster – now it’s time to practice your delivery. Consider your audience...details or omit crucial background. Practice a few times while standing at your poster – it’s a different...unfamiliar venue. Finding your poster board well ahead of time ensures you aren’t scrambling to tack up your poster...before the session begins. Plus, you’ll have extra time to practice at the actual venue. Introduce yourself...to a poster where the presenter spends the entire time talking to the poster, as opposed to engaging with...on the recent literature in your field. Take your time: We often get so nervous when asked a question that...

...Awardees: Xin Zhou and Fatima Enam. There were many excellent applicants but Xin and Fatima stood out for their...our recent blog post. Fatima Enam: Biosensors for human milk oligosaccharides Fatima Enam is currently a...beneficial bacteria in the newborn gut. One of Fatima’s ultimate goals is to engineer bacteria for the efficient...technologies are too time consuming in the first case and nonspecific in the second. Fatima therefore set out...discoveries. Read on to learn more about Xin and Fatima's work and keep checking back in with the Addgene...Light-inducible Proteins, or FLiPs.  Xin later optimized these first generation FLiPs by mutating Dronpa...healthy microbiome. In order to achieve this goal, Fatima first needed an efficient and cost-effective way...

...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...