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...for some time now. When titrating AAV by qPCR, viral DNA is amplified and monitored in real time. When the...s. Increasing the elongation time to 2 minutes and the denaturation time to 1 minute has been shown to.... But what if we could repeat this assay 10,000 times to ensure that our titers were as accurate as possible... or a reference, meaning saving on reagents (and time)! AAV titering using droplet digital PCR Droplet... that could potentially lead to contamination. Optimize your PCR For a good starting place on what PCR...

...can be shortened since the plated cells will have time to produce beta-lactamase prior to division, instead...effects of the antibiotic. This buys you a little more time, since bacteria won’t divide immediately after it...beta-lactamase. Pros: Widely available, cost-effective, timesaving for transformations. Cons: Prone to satellite... Pros: Very stable, interchangeable with Amp, timesaving for transformations. Con: Expensive.   Kanamycin...resistance Choosing the best antibiotic resistance ultimately comes down to your applications and needs. The...selecting!   Antibiotic Resistance Stability Time Saving?  Works for multiple organisms? Compatible...storage temperature and pH on the stability of antimicrobial agents in MIC trays. Journal of Clinical Microbiology...

...neuroscience, including transgenic mice, neuron-optimized viral vectors, and high-throughput screening approaches...Figure 1b), and sgRNAs can be found here. Neuron-optimized viral vectors Alternatively, Cas9 activators can...recently by the Day Lab, who developed neuron-optimized lentiviral vectors for in vitro and in vivo CRISPRa...with BioRender.com. The Day Lab used their optimized vectors to highlight a key advantage of CRISPRa... to neuronal physiology. The Day Lab’s neuron-optimized lentiviral vectors can be found here. High-throughput...not all neuroscience applications require such optimized tools. This is elegantly demonstrated by the Qi...PMC6214761. Savell, Katherine E., et al. "A neuron-optimized CRISPR/dCas9 activation system for robust and ...

...your screen result may or may not actually help. Ultimately, if you cannot describe the likely outcome with...grant. Either way, think realistically about the time and resources required to feasibly accomplish the... address the aims in more depth in the remaining time. The typical chalk talk structure is: What is the... details. Make the most of your limited speaking time. Beyond the information you seek to convey in your...feedback. It’s also helpful to practice at least a few times with people that are outside your lab and outside...or while the committee is arriving. Spend as much time as possible facing the committee and not the whiteboard...secretory proteins in the Endoplasmic Reticulum, optimization of fluorescent proteins,  and live cell imaging...

...us, such as finding time to read all of these articles. Most of us hardly have time to keep up with all... our own topic. Only on occasion do we then have time to read about research from other disciplines, or... of interdisciplinary research while at the same time increasing interdisciplinary citations in their ...videos of 3 to 5 minutes provide more than enough time for researchers to describe their science, and for... Rely on Circadian Rhythm and Melatonin for the Timing of Nocturnal Courtship Vocalization. The video ...youtu.be/EHHhiawIdWM Saving salmon, one embryo at a time – A video by LeeAnne Thayer, a Ph.D. candidate in...

... gene you’d like to manipulate in mind. Now it’s time to consider where you would like to attack this ..., 2014. Not all gRNA’s are effective. At the time of writing many of the specifics surrounding why... remain unknown, but fret not! The key to saving time and energy during your in vivo work is to screen...similar plasmids for gRNA expression), one gRNA at a time. After 48 hours of incubation, you assess the level...technique, you can assess several sgRNA’s at the same time, and choose the most effective ones to increase ...validated your gRNAs for use in mammalian cells, it’s time to use them in mouse embryos. When researchers first...mouse embryos to develop. Once they are born, it’s time to screen them for your desired edit. Good news!...

..., in actuality, many find using lentivirus to be time consuming, difficult, and lacking in reproducibility...enough to promote cell growth. Overgrown cultures stimulate quiescence, the resting phase of the cell cycle...efficiencies. Cultures should only be passaged 20-30 times before discarding and thawing a fresh vial. In many...prefer to collect and pool the harvests from multiple time points while others opt for a single harvest after...lentivirus are determined by the envelop protein. Many times lentiviral vectors are engineered with non-native...such variables as cell density, volume, incubation time, and MOI can all impact the success of your lentiviral... S., Guo, S., Xu, J., Kappes, J.C., Hel, Z. “Optimization of the transductional efficiency of lentiviral...

...is that access of this data is to be made at the time of manuscript submission. The rationale includes...these were those from researchers who wanted more time to mine their own data, with some smaller labs feeling... originator was oblivious to, no matter how much time they had. It could be argued that data generators...having exclusive access to data and enjoy reasonable time to make sense of it, with their decision to publish... specific (expensive) software that evolves over time such that prior formats are no longer supported/... especially for young researchers who may be intimidated by having to contact more senior colleagues. ... kinases implicated in human diseases. His spare time is spent trying to raise the alpha isoform of GSK...

...this time, Dr. Koeris and Professor Timothy Lu (a graduate student in the Collins' lab at the time) got.... As you'll hear Dr. Koeris explain however, the time was not right for the development of phage-based...listening! Download the podcast here! Podcast breakdown (Time : Topic) 0:00 - 1:25 : Introduction 1:25 - 6:19 ...   We'd like to thank Dr. Koeris for taking the time to talk to us and taking up a position on the Addgene...

...transplant — creating intense time pressure for the research team. Ultimately, the lifesaving effort was ...contribute to more in the future. Figure 1: A timeline of CRISPR base editing advances that led to KJ...indel mutations that plagued CRISPR editing at the time. KJ was born with a C→T mutation on one allele of... of off-target editing and longer genome search times. Using a machine learning approach, the lab proposed...patients. While these gRNAs would still need to be optimized and assessed for off-target risks, such a modular...development of genetic therapies even further. In the meantime, base editing will undergo even more improvements...

...interest as possible, some optimization will be necessary. To begin, one should optimize protein production using...phosphoserine directly into recombinant proteins (5). Optimization of this system defined three components required...phosphoserine, the Sep-tRNA synthetase, and EF-Sep – an optimized elongation factor that accepts Sep-charged tRNAs... have worked most extensively worked with and optimized Sep-OTSλ and can recommend its use (6).  2. C321...that can be used as a positive control during optimization experiments. This version of GFP will contain...purification strategies; expression should be optimized for parameters such as induction temperature and...

...develop an optimized photoactivatable CRISPR gene activation system, but settled on a different optimal fusion...to use light to externally control the location, timing, and reversibility of the genome editing process...researchers tested a variety of combinations to optimize both fusion proteins. The best performing combination...fusion protein combination. The optimized LACE system consisted of: 1) CIBN-dCas9-CIBN, where CIBN is the... prevent DNA binding in the dark. Upon light stimulation, the pdDronpa moieties move apart, allowing DNA...LOV2 has a C-terminal helix that unfolds when stimulated by blue light. When fused to the catalytic site...chromatin architecture, it is useful to regulate the time in which dCas9 binding occurs. Bubeck et al. expressed...

...continued this tradition. We now have over 30 full-time employees, so we created one long lunch table and...bring in baked goods and we have a few unique lunchtime traditions that might be fun to incorporate into... take a break from their everyday tasks and make time for special projects? We borrowed an idea from the... success at accomplishing “we-should-really-make-time-for-that” type projects. Cross-team “Random Lunches...between labs. Scientists are often willing to make time for free food, and these lunches would provide a... as important to take a break from work to share time with the people around us. It makes the quality ...

...nucleome, the organization of the nucleus in space and time, and to understanding how nuclear organization varies... change in response to the cell cycle or other stimuli.   To create a colorful Cas9 toolbox, Ma et al....study is the method’s sensitivity. The authors estimate that a minimum of 150-200 fluorescent protein ...phototoxicity concerns and extending the potential imaging time for this technique. The NIH Common Fund has made...epigenetics and cellular reactions to external stimuli. It could also have major applications to cancer...imaging of genomic loci in living human cells by an optimized CRISPR/Cas system. Chen B, Gilbert LA, Cimini ...

...more practical lab experience to later gain a full time position in a lab. I worked on the genetics of Wilms...of interest before you dive straight into a full-time position. For my part, I really enjoyed some aspects...the company was to fulfill contracts, my work sometimes felt rushed and the focus always seemed more on...has been rapidly growing. These factors left me optimistic that I would find a genetics related role soon...situation, communication between departments was sometimes difficult and the solution attempted was more ...solved with good management practices, but can sometimes go unnoticed at large organizations. Another downside...because you need to work independently most of the time and plan your own work schedule.  I really love ...

...during this time more visible and thereby have a positive impact on science. ReFigure saves time, knowledge...continue reading for more details. How much time do you spend reading scientific literature? Very...experimental results but, obviously, this is not optimal. What if you could save the figures of interest... instantly and are editable by the author at any time. Insights contained in your ReFigure could be beneficial... Resources on the Addgene Blog Use Apps to Save Time and Track Your Experiments Use video to share your...

...videos is very time consuming; producing a 1-2 minute video takes an entire week of our free time. After taking...preconceived obstacles, optimistically grasp at limitless potential, and who sometimes wish they could live...about anything can disrupt your bench work and sometimes no culprit can be found…. In our lab we call this...practically universally loved (except for those rare times that we step on them with bare feet), to connect...

...s Lab (plasmid page here). Now I spend much less time wrangling bacteria and instead help Addgene fulfill...Addgene? So why did I join Addgene? Was I jaded by my time in academia? Did I run screaming from the lab like...Outreach Scientist where I spend a large amount of my time meeting with and talking to graduate students, post-docs...teenage years I was super shy - there was even a time when I was too nervous to ask store clerks for help...graduate work at Harvard, I recognized that I would ultimately want to go into science communication but thought...writing, teaching, presenting, analytical thinking, time management, information distillation, etc) are transferable...

... other reagents. Virus distribution came up time and time again with many researchers lamenting the viral...Instead of spending time and money producing virus from select vectors in the repository yourself, you...frustrating process that requires a large investment of time and resources for labs that don’t have experience...titering, and testing viral preps for them. At the same time, we’re developing ourselves as a knowledge base ...to be distributing new types of materials. At the time of publication we’ve already fulfilled more than...

... Working in a lab is nothing like a typical part-time or 9-to-5 job. I had to ask to find out what a “...Professor, Assistant Professor, Associate Professor Sometimes teaches graduate and/or undergraduate classes ...that not all advice is good advice, and it is ultimately up to us to decide whether we want to take it...mentor and choosing the right lab. Remember that your time is valuable! Don’t jump on the first opportunity...whether it’s something you want to dedicate your time and energy to. How can I make the most of my experience...to ask “dumb questions” and to make mistakes. Ultimately, though, scientists are all just people. I’ve...cultivate stronger relationships with my labmates. Sometimes what I thought was a dumb and very basic question...