We narrowed to 867 results for: PLE

...downregulate multiple genes at once. Read the paper in mBio Find the CRISPRi plasmids A multiplexed gRNA piggyBac...Base editing CRISPRi in Caulobacter crescentus Multiplexed gRNA for human pluripotent stem cells CRISPR-... Blue light renders the NLS accessible and the complex enters the nucleus where dCas9-Mxi1 can repress.... In the dark, the NLS is inaccessible and the complex remains in the cytoplasm. Read the paper in Cellular...differentiation, but it takes several weeks to delete or deplete individual genes. Michael Laub’s lab developed ... deliver uniform and sustained expression of multiplexed gRNAs in human pluripotent stem cells. Previously... construct and the fluorescently-labeled gRNA multiplex. Read the paper in Scientific Reports Find the...

...field. Fundamental principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores...attached to self-labeling proteins. Some common examples of these self-labeling proteins are SNAP-tag® ...of protein substrates/peptides and antibodies. Example applications of this include coupling to an antibody...specific receptors is key, especially for G-protein-coupled receptors (GPCRs). Mutations in receptor sites ...photons absorbed over the photons emitted. For example, DAPI is much dimmer than Alexa Fluor 488, and ...signal and facilitate clear differentiation between multiple dyes during analysis. Image resolution is impacted...microscopy techniques, such as Stimulated Emission Depletion (STED), and Stochastic Optical Reconstruction ...

...there are some genetic factors involved. For example, multiple studies show that certain mutations of the...is a terrible disease that is, unfortunately, completely preventable. Mainly caused by exposure to asbestos...mesothelioma instead rely upon private donations. Many people think that because mesothelioma is so rare, there... researchers don’t entirely understand why some people develop the disease while others never do, but ...slow the growth of tumors. More often than not, multiple techniques are used as part of a treatment plan...positive potential for treating mesothelioma. For example, patients who have peritoneal mesothelioma – that... be used to distinguish between three groups of people:  Those who have mesothelioma Those who have been...

...fields in order to provide useful solutions to complex challenges. As a cross-disciplinary researcher,...Many of the challenges in science today are too complex for any single discipline to solve them on their...interdisciplinarity index of research for single or multiple disciplines – you can select the disciplines and...few years, I decided to look at what online tools people generally like to use to share ideas – with the... in terms of communicating ideas. In 2012, for example, 3 billion hours of video were watched each month... disciplines leading the way, the future of complementing published research with video overviews was ...networking, visual appeal, instant sharing across multiple other sites), I created a new platform for sharing...

...approaches that may come up during discussion.  One example of agility at work is our process for working on...instructions on how to demonstrate the feature. For example, we recently reduced the number of steps involved...stories in that sprint relate to the theme. For example, we could have a shipping sprint and all the stories...get different perspectives on how things are implemented and from that, develop better software.  Maintenance...how you combine it with other ingredients.  For example, we use a third party library to handle label printing... compatible with the updated library. A recent example is updating our software from Python 2.7 to Python...analogy. Human languages evolve over time, for example from Ancient Greek to Modern Greek. Books written...

...stimuli. One way they do so is through G protein-coupled receptors (GPCRs), receptors that play many roles...GPCRs (Takeda S et al., 2002) and many downstream complexes involved, it can be difficult for scientists to...in sight (Olsen RHJ et al., 2020). A transducer complex consists of a trimer of three subunits: alpha, ...collectively known as a heterotrimeric G protein. This complex converts a signal received by a GPCR on the cell...is being able to identify specific transducer complexes that are activated in response to a ligand binding...inactive, these subunits are found in a trimeric complex bound to the intracellular surface of the GPCR....Following activation in response to the ligand, the complex will disassociate, resulting in a decreased GFP2...

...career Developmental biology caught me early on: a simple egg is packed with information. I am fascinated...individual diversity to get from gamete to a visibly complex - and unique - adult. My initial research directions...teacher was looking for an outlet. I started asking people: why do you do what you do? How do you like it?...it? how did you get there? Many of those people I met through AWIS (Association for Women in Science); ...visiting seminar speakers. My eye was caught by a couple of unusual job openings, although they seemed too...applied research. I focus in general terms on a couple of very broad topic domains, but each day brings...and surprisingly diverse manuscripts. I work with people, nurturing networks, as much as with technical ...

...adopting simple note-taking tools, others using excel sheets or organizing folders in complex ways, there...lengths to keep accurate records, but given the complexities of modern research, it’s often not enough. A...if they are to switch from paper to digital for example): “… 99% of respondents indicated that ease of ...developed the SciNote Electronic Lab Notebook. Implementing an electronic lab notebook starts with defining...property across various software providers used to supplement their paper notebooks. Instead, all of the important... you’ll be able to do it when needed. How many people from your lab would use the ELN i.e. do you need... facilitate this, some ELNs allow you to invite people and assign them roles and permission that only ...

...scientists gather information about reality informing people and their governments on what outcomes are actually...earlier analogy, no amount of shouting will make an apple fall up. As scientist and politician Rush Holt said...and public opinion do not seem to take note. For example, 91% of active research scientists (scientists ...consequences While frustration and anxiety have inspired people to rally for science, others are concerned about... is not going change depending on the number of people supporting it.  3) Linking scientists to one particular...march. Who is marching and why? Who? All sorts of people are choosing to march on April 22nd. As of April...for science education and literacy as I believe people that can see and understand data themselves can...

...proxy for dimerization (Shaner et al., 2008 - supplementary figure C). However, these assays are qualitative...fluorescent protein. These were either (i) coded by the multiple cloning site and therefore consist of random amino...protein is cleaved by proteases. Nowadays, with ample structural information available, it’s clear that...protein of interest (POI). To indicate the sequence, people often use terminology like “C-terminal fusion,”...#85045). We often use restriction enzymes in the multiple cloning site for generating fusions. For the POI-fluorescent... acids (PVAT, when we use the AgeI site in the multiple cloning site) is sufficient. For the fluorescent... better preserves the function of the POI. For example, the APT1-mVenus fusion localizes at the Golgi,...

...pri-miRNA step. These RNA duplexes bind to the RNA-induced silencing complex (RISC). After binding to ... You need to include both sides of the complementary RNA duplex and the hairpin loop sequence (Figure ... of hairpin structures to create a mature siRNA duplex. Fun fact! Endogenous miRNA production starts with...create pri-miRNA, which is then processed by a complex called Drosha to pre-miRNA. miRNA plasmids are ...to RISC, the RNA duplex is processed into a single targeting strand, with the extra strand usually being...typically range from 19–22 nucleotides and are 100% complementary to their target sequence. However, you can’t...action, and degradation of RNA-induced silencing complex. Molecular Cell, 82(1), 30–43. https://doi.org/...

...abundance, consider the following example gel. Figure 5: An example gel (left) with bands corresponding...western blot requires a standard curve with a pure sample of your protein of interest (POI). This approach... a perfect world, protein concentration in your sample would directly correlate to protein abundance on...calculate things about earlier ones. In practice, multiple sources of error (red) influence these relationships...not perturbed by the experiment Unfortunately, multiple reports suggest that common housekeeping proteins...are highly expressed in cells. This means that at sample concentrations appropriate to detect a less-abundant...be measured in your assay. This is influenced by sample concentration as well as by antibody dilution —...

...lympho-/splenocytes, you might only be interested in one of the various cell populations; for example, CD8...These methods can include labeling markers with multiple proteins/antibodies, targeting markers found within...beyond standard surface labeling. Pro tip! Many people use 'labeling' and 'staining' interchangeably to...antibody can originate from a range of species; for example, rabbit or mouse. In a second step, a fluorophore-conjugated...Otherwise, your secondary antibody will bind to multiple primary antibodies, yielding a false positive ...FACS achieves greater purity of your sorted cell sample than MACS. An effective strategy to sort out your...the permeabilization solution can be the same). Examples of such targets include transcription factors,...

...avidin can be added to your sample. Any free biotin-binding sites on this complex can bind to your biotinylated... proteome that are biotinylated. In E.coli for example, the BPL, BirA, is responsible for biotinylating...plague these animals even if their diet was being supplemented with biotin. Thus something in the egg white...discovered in the 1960s, strepavidin. The Biotin-avidin complex can then be used and/or manipulated for a scientist...immunoprecipitation experiments. Biotinylated proteins or complexes can be easily purified using columns or beads ... added to the reaction for easy detection. The multiple biotin binding sites found on avidin are also ...This creates a large meshwork of biotin-avidin complexes bound to your protein of interest leading to amplification...

...SciComm skills and I had to coordinate with other people for a variety of projects. I was therefore forced...its utility) How much time it will take you to complete the task When picking which tasks to actually...accomplished quickly and things that will take longer. Completing the quick and useful tasks will swiftly give ... accomplishments will likely energize you. For example, let's say you’re doing a multi-day experiment ...experiment. The first way, it will take you 3 days to complete the entire experiment, but you’ll have to spend...experiment. The second way, it will take you 5 days to complete the experiment, but you’ll only spend half of ...remember things better when I write them down. Some people also find it useful to write out their to-do lists...

...equipment, reagents, animals, glassware, and even people. When developing a budget, I suggest that you focus...consumables (such as pipet tips, glassware, etc), get a simple list from your current P.I., lab manager, or administrator...need to tap for your future research team. For example, if you’re going to need a tech because you’ll ...regions where the host institute is located. For example, salaries in Kansas versus salaries in NYC will... that have the means to support your work. For example, if your work requires specialized equipment, try...need.  Prioritizing resources in your budget The simple practice of putting together a resource budget ...rather than focusing on long term investments. For example, purchasing “used” equipment can save you money...

... shape what people know and believe about the world. It is vital to understand what people know – or, ...like?  One of my more recent studies looked at how people perceive and evaluate peer-reviewed sources of ...called the “biased evaluation of evidence,” in which people (conscious or unconsciously) pick apart sources... to dismiss them. There’s evidence to show that people with greater levels of science literacy are especially...hands-on science communication. Between New Zealand’s multiple Covid-19 lockdowns, I held mask-making workshops...the difference between airborne and aerosolised droplet transmission and how different fabrics and mask...illness. I later helped run workshops where I showed people how they can make reusable cloth menstrual pads...

...minipreps for small batches of samples. As soon as that process is complete, QC scientists prepare Sanger...mutation.  Once the samples arrive, each group of plates will have select samples prepared and screened...collection in your experiments and published your data, please fill out this form and we’ll add your citation ...available. Plasmids are continually added as they complete QC and you can find all available plasmids listed... high copy backbone with a T7 promoter driving multiple combinations of WT, codon optimized, untagged,...screened by NGS to spot check the samples, the rest will be put into a status we call “Archived.” This status...for all several thousand plasmids at once. Any samples that are requested by scientists, will immediately...

...you need more than one?   Multiplex assays Multiplex assays involve multiple antibodies, each with a different... the antibodies time-sensitive, using precious samples, or being compared to other data in a manner that...this post, the conjugates, or signaling molecules! Please keep in mind that this is just a brief introduction...conjugate that can be activated to emit light with a simple kit, and is readily available on any number of ...ain’t broke, don’t fix it. HRP is easy to find, simple to use, not sensitive to light in its inactive ...detected in the same assay. This is one of the most complex subjects in antibody-based protocols - but thankfully... this is an Antibody 101 post, so we'll keep it simple. If you follow the two basic guidelines below, ...

...cerevisiae yes - This can complement ura4- S. pombe, but the complementation is weak. yes   LYS2 L-lysine...no S. cerevisiae yes - This can complement leu1- S. pombe, but multiple copies are required. yes   TRP1...vectors? Yeast are eukaryotes and thus contain complex internal cell structures similar to those of plants...mechanism of homologous recombination to facilitate simple gene replacement/mutation. Yeast expression plasmids...selection markers.  The yeast origin of replication (Please note: This first section primarily pertains to ... including a copy of a functional gene which complements the host’s auxotrophy. When grown on media NOT...drug selection methods feasible in yeast as a complement or alternative to using auxotrophic markers. ...