We narrowed to 867 results for: PLE

...when studying them since their complete knockdown is lethal. Incomplete or titratable knockdown, however...the tools scientists can use to study them. For example, gene knockdown with CRISPR interference (CRISPRi...wall. Once in the recipient cell, the CRISPRi DNA (purple) flanked by left and right Tn7 end sequences (green...showed that Mobile-CRISPRi is transferable into multiple bacterial species including several involved in...pathogenic strains and environmental isolates. For example, Peters and colleagues used Mobile-CRISPRi to knockdown...if that gene is a target of an antibiotic. For example, let’s take a look at the essential gene folA which...also possible with Mobile-CRISPRi. As a proof of principle, Peters and colleagues constructed a 40-member...

...increases. Multiple inserts: There are some cloning methods that make working with multiple inserts a ...as some methods are more amenable to combining multiple DNA fragments than others. In today’s post, we...with a special emphasis on these five categories. Please note that many of the following cloning methodologies...interested in learning more about any one of them, please click on the links throughout the text or see the...per reaction but can be used to quickly generate multiple clones. Difficulty: If you’re an experienced cloner...Golden Gate assembly allows one to simultaneously complete an ordered set of scarless ligations following...one right answer, so don’t be afraid to explore multiple approaches to find out which one works best for...

...into multiple cell types of the same lineage. Blood (hematopoietic) stem cells are a good example of multipotent...mostly to maintain cell numbers or replenish dying cells. For example, hematopoietic stem cells found in...functions [1]. In many tissues, stem cells serve to replenish/replace damaged cells that no longer function ...[1]. Stem cells’ ability to differentiate into multiple cell types makes them useful models for developmental...thought of as descendants of totipotent cells. An example of pluripotent cells is embryonic stem cells (ESCs...in science communication and publishing after completing her postdoctoral training from Massachusetts ...

...yield. Keep in mind that you might need to use multiple techniques for a particularly stubborn prep. ...remove endonucleases (see figure at right for an example). Look out for carbohydrates Certain strains, including...the cell lysis process. To ensure that lysis is complete, use a reagent like Qiagen’s LyseBlue, which uses... uses color to show you if lysis is incomplete. Protein expression strains are not good for cloning Bacterial...yields. Why would volume make a difference? In the simplest of terms, more volume = more cells = more plasmid...culture media Changing to a richer media or adding supplements can improve your cell density, leading to an ...normally recommended antibiotic concentration. Add supplements to the media You can achieve higher cell density...

...cameras with electron multiplying couple-charged device (EMCCD) or complementary metal-oxide semiconductor (...neurosciences.  These developments in microscopy coupled with techniques that make tissues transparent are...beam into a thin sheet to illuminate a fluorescent sample and captured the reflected light using a different...combined light sheet microscopy with sequential multiple-view reconstruction (5). In order to image large...microns, illuminates only one thin plane of the sample surrounding the focal plane of the detection lens...problem is to illuminate and acquire images of a sample from two opposing sides and merge the images, such...through Addgene.  The combination of LSFM with multiple tissue clearing strategies and neuronal labeling...

...ligands to the same sample to label sub-populations of your protein of interest. For example, the Cohen lab...derivatives of its substrate O6-benzylguanine (BG) (Keppler et al., 2003). This engineered hAGT later became...multi-component systems can be complicated to implement, they can be highly specific with a small and ...Atto590-BG (green), imaged with stimulated emission depletion (STED) nanoscopy. Scale bar, 2 μm. Inset shows...proteins in vitro, you can even use the same tag for multiple proteins but label each separately before reconstituting... assembly-regulating proteins in vitro reveals complex events. a) Micrographs of a fluorescently-labeled..., biotin, or magnetic beads. In one innovative example, the Zhang lab used SNAP-tag to decorate synthetic...

... disturbing trend in the number of high profile people denying science, from celebrities falsely claiming...scientists questions about their work. Thousands of people showed up; some with some really great signs. The...work because it's important to our health.  Most people won't argue with that idea, but they might feel...figures.  Ask me about my science!", read my sign.  People asked, and we talked.  That's a win. Mark Lucera..., challenged us. I worried the march would be a simple, unquestioning celebration of science, at a time...funding, and teaching experience that it takes to complete your resume, without ever thinking that research...things about science. We need to keep encouraging people to look for evidence through education.  My only...

...When Karl Barry Sharpless was awarded this year’s Nobel Prize in Chemistry, he joined an elite club, ...his program. Initially, Provost made his labs completely open-ended, allowing every student to work on... the culmination of many such experiences. For example, students in Provost’s course recently used a combination...Kowalski et al., 2016; Scott, 2021). In fact, completion of Provost’s CURE increased students’ confidence...Community (MCC) that seeks to reduce barriers to implementation. Funded by the NSF (with Co-PIs Jessica Bell...for your classroom, I highly recommend you try implementing one. It's likely only a matter of time before...Poirier, V., Pellettieri, J. 2020. The exon junction complex is required for stem and progenitor cell maintenance...

...removing excess dye from the sample, which is particularly important for samples where washing is difficult...reason is something called “the photon budget”: each sample has a limited number of fluorophores and each fluorophore...of information one can extract from a biological sample is wholly dependent on the photon budget and pushing...fluorescence microscopy often requires more photons. For example, moving from transient overexpression of protein... desperate to extract a few more photons from a sample. Chemical fluorophores can be substantially brighter...labeling chemistry easier and more functional in complex biological environments such as live cells and ...brightness of the resulting conjugate, and the complexity of the system. A particularly desirable property...

... for example, helping to improve the papers submitted to journals. In addition, it is my pleasure to interact...enChIP system enables us to express the CRISPR complex much easier in various cells, including primary...technologies consisting of iChIP and enChIP are recent examples. We also developed the inducible translocation...and PLoS One. Additionally, you are involved in multiple professional scientific societies. Why is being...system of journals and grant applications, for example. However, at the same time, it's fun to be involved...give to post-docs and graduate students as they complete their appointments in your lab?  Fujii: These ...develop new technologies, hopefully they'll be completely different from what we are now doing in my lab...

...and replaced it with a double-bonded oxygen. This simple enzymatic reaction enabled conversion between the...and one-ring pyrimidine base. This is a far more complex chemical task, not a job for a single enzyme. ...bind DNA, Cas9 requires (1) a guide RNA, (2) a complementary DNA sequence, and (3) a protospacer-adjacent...that site is filled depends on cellular factors. Multiple labs tried to identify beneficial factors to include...separate and preventing repair based on the complementary strand. This could explain why the edited U ...original G is not primarily converted back to a complementary C. Optimized base editing with eOPTI-CGBE and...so the Yidi Sun and Erwei Zuo labs tested and implemented several of these advances for CGBEs (Yuan et ...

...reprogramming factors can be delivered and expressed in multiple somatic cells via various methods. Some of the...directed differentiation.” Specific cell media, supplements, bioactive small molecules, and growth factors...natural transdifferentiation is rare in mammals, an example is observed in the pancreas when excess β-cell ...1987, Davis et al. reported one of the earliest examples of transdifferentiation in vitro where treatment... of interest for in vivo transdifferentiation. Multiple cell types such as fibroblasts, hepatocytes, and...vitro Both in vitro and in vivo Epigenetic marks Complete removal to convert to iPSCs and re-establishment...major processes by which cells are converted to replenish and repair cells that are lost or damaged. Both...

... The central nervous system (CNS) orchestrates complex processes enabling organisms to control their movements...the visible spectrum, researchers can also use multiple FPs in single experiments and visualize numerous.... smFPs are literally composed of FPs fused to multiple epitope tags. Indeed, to create the smFPs Looger...labeling of these fine structures through their multiple, high-affinity binding sites for primary antibodies...expressed and/or similar proteins Through their multiple, highly specific antibody binding sites and primary...used to distinguish highly similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell...super-resolution STORM imaging, and electron microscopy. Sample preparation for these techniques sometimes makes...

...specific examples. Instead of saying “I have excellent communication skills” try giving an example, like ...application process, it's a good idea to talk to people in the field to understand what the norms are. ...match the job requirements, by giving specific examples of your work and explaining how your experience...pointing to your transferable skills. In the above example, if you don’t have experience with ddPCR, but do...beginning is often the hardest part, here are some examples to get you started! Dear Dr./Professor [last ...strengthened an application. Even so, there were examples of great hires who hadn’t submitted a cover letter... Multiple Authors...

...certain loci to create changes in phenotypes. For example, CRISPR activation has been used to differentiate...gRNAs targeting every gene in the human genome for example, can be made easily and cheaply using oligo library...allows for exploratory-based research as well. For example, CRISPRa can be used to find potential cancer drug...et al., 2019). Because of its ability to target multiple gene loci simultaneously, CRISPRa can also be ...(Chavez et al., 2015)) are good choices across multiple cell lines (HEK293T, MCF7, U2-OS, Hela, N2A, 3T3...Church G (2016) Comparison of Cas9 activators in multiple species. Nat Methods 13:563–567 . https://doi....transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https://doi.org/10.1038/nature14136...

...of storage. For example, mouse behavioral geneticists may track the motion of multiple animals on different...limiting numbers of figures such that many have multiple, complex panels. PLOS, itself, has not finalized its...Sharing Policies, etc.) and is founded on the principle that scientific publications must contain enough...some data, etc.  The firestorm that followed implementation of the policy comprised of numerous objections... paper are in scope. For images, the number of examples supporting the published material is a factor ...collections and interpretation of data, publishing complete data, rather than just a single projection of ...

...Benjamin Franklin Effect, most people actually enjoy helping (just ignore the people too busy to respond to you...can do during your training is meet interesting people doing interesting things. I call this building ...insights you will get from the interaction. Most people like to help One good place to look for interesting...Sharing a school is a little connection that many people honor. Talking with strangers is usually challenging... with strangers. Be bold in asking all kinds of people to connect with you and to share their stories....uncover interesting opportunities by meeting with people working in diverse and unusual fields. My favorite...

...out of protein purification through simple and efficient multiplexing. pCri plasmids are divided into two...schematic below), enabling a researcher to test multiple tags for one protein quickly and easily. These...one need to remove the tag post-purification. A sample of pCri constructs: pCri-1a, -4a and -9a. 6xHis...on these and other tags used in the pCri system, please see Plasmids 101: Protein Tags. Beyond E. coli...purification. For a more thorough introduction to pCri, please see the original publication cited below. pCri ...plasmids are available either individually or as a complete kit from Addgene. References 1. Goulas, Theodoros...

...to the popularity of GFP including (i) fast and complete maturation to functional, fluorescent protein ...endogenous fluorescent co-factors can be used. Examples include the LOV domains that bind flavin (Buckley...under acidic conditions, with pKa values below 4. Examples include mTurquoise2 (pKa = 3.1), tagRFP (pKa =...diameter of about 2.4 nm (Hink et al., 2000). The complete beta-barrel is necessary for its fluorescence ...specifically and with high affinity to fluorophores. Examples are the FlAsH tag, the His tag and several others... bind the molecule of interest. For instance, pleckstrin homology domains can be used to detect specific... proteins are also a better choice for thicker samples. Another solution to avoid autofluorescence is ...

...recombination intermediates can then resolve to complete the DNA repair process Homology directed repair...happen on the crossing strand (horizontally at the purple arrows) or on the non-crossing strand (vertically...is elongated and ligated to fill the gap, thus completing SDSA.   Break-induced repair (BIR) pathway BIR... and even easy. This targeted-break technology coupled with HDR is a powerful site-specific knock-in tool... uses reverse transcriptase to generate the complementary ssDNA. Easi-CRISPR was shown to increasing editing...series was designed to help explain the basic principles driving CRISPRs and genome editing technology...Pfeiffer P, et al. Mutagenesis (2000). PubMed. Multiple Pathways of Recombination Induced by Double-Strand...