...achieved by simply replacing the product in question. This can solve the problem in a simple way even if...(e.g. a postgraduate degree in law, an MBA, or similar), there are many roles out there that require exactly...Technical Support Advisor, Technical Correspondent or similar. While there isn’t a clear distinction between ...

...A similar genetic code is used by most organisms on Earth, but different organisms have different preferences...amino acids (see example to the right). With such simple rules, you might think it’s easy to come up with... express proteins in heterologous hosts. If you simply amplify a gene of interest from the human genome.... This is known as “codon optimization.” Though simple in theory, this is not as easy as it sounds. Even...

...oligo library synthesis. Prior to Cas9 activators, similar tools were made using other DNA binding proteins...change the sequence targeted by the activator by simply providing a new gRNA rather than engineering an...of its ability to target multiple gene loci simultaneously, CRISPRa can also be used to map molecular ...given cell type or organism, have been used in a similar manner to Cas9 activators. However, these can be...

...is completely deleted (designated lon or Δlon) similary reduce proteolysis of the expressed proteins. ... can neither restrict nor methylate DNA. dcm: Similarly, strains with this mutation are unable to methylate...coli T5 promoter (found on vectors such as pQE or similar), inducible under IPTG (lac repressor on the pREP4...variety of sources, so fear not—there are a few simple troubleshooting measures that can help get you ...

...technique, but it is very important to follow some simple rules to avoid common pitfalls and to use it at...recommend to using two fluorophores that have a similar brightness - even though in the most popular pair...most efficient approach for your own system. The simplest and the most popular one is the sensitised emission...photobleaching method The acceptor photobleaching method is simple but limited to a single measurement. This method...

...models due to their size, physiology, and genetic similarities with humans. However, rats have some key differences...axolotl may reveal the potential humans have for a similar degree of regeneration.  In addition, axolotls ...have many age-related phenotypic changes that are similar to those observed in humans. Thus, understanding...them an ideal model organism; they are relatively simple to maintain and breed and they have large, easily...

... E, de Simone S, Colella P, Iodice C, Polishchuk EV, Sommella A, Colecchi L, Rossi S, Simonelli F, Giunti...in mice. This vector transduced ~10% of cells, similar to the transduction rate of a single AAV. Odom ...Trapani I, Colella P, Sommella A, Iodice C, Cesi G, Simone S, Marrocco E, Rossi S, Giunti M, Palfi A, Farrar...

...techniques and strategies one can employ beyond simply incubating cells with single-fluorophore-conjugated... B cells, and dendritic cells. In this case, we simply need to label the lineage markers of all cell types...selected color will not be CD8+ T cells and we can simply “dump” them by gating on the negative population... new methods of target visualization beyond the simple surface labeling. Let’s briefly walk through the...

... to a Perfect Science Talk." 4. The chalk talk, simple in principle The first question I usually get about...focus of your first grant? (1-2 sentences) Simply stated, what are your aims? You could directly ...official aims, but I suggest distilling them into simple goals/outcomes. This isn't the time to go into ...in the knowledge that you have given everyone a simple plain language map of where you are going and what...

...trace file generated in Sanger sequencing and a simple algorithm to quantify editing efficiency by insertion...unedited population and your Cas9-targeted cells. Simply upload both trace files to the TIDE tool online...knock-ins both large and small is a software very similar to TIDE known as TIDER (Tracking of insertions,...main advantage of NGS is that it allows you to simultaneously validate edits and look for off-target changes...

...that strongly binds the glycoprotein, avidin or a similar protein, streptavidin. This strong interaction ...Thus to capture protein complexes, scientists can simply express a bait protein fused to BirA* in cells ...identify additional interacting proteins in one simple experiment. PCA is an in vivo technique where two... and Paul A. Khavari. "Research Techniques Made Simple: Emerging Methods to Elucidate Protein Interactions...

...next-generation sequencing techniques to answer similar experimental questions (see below). Importantly...location of a given nucleotide, a researcher (or a simple computer program) can then reproduce the DNA sequence...generation DNA sequencing machines (9). Today, similar technologies, sometimes referred to as sequence...Kathleen, and Daniel Nathans. "Specific cleavage of simian virus 40 DNA by restriction endonuclease of Hemophilus...

...cassette (Dickinson et al., 2015) made this step as simple as heat-shocking to induce the expression of a ...my initial direct screening efforts, which were similar to the approach detailed by Paix et al. Although...Addgene. In 2019, Farboud et al. described another similar co-conversion marker (repair of a zen-4(cle10ts...choosing. Tools such as SapTrap Builder greatly simplifies design of these constructs, allowing novice researchers...

...reason most people enter postdoctoral training. Similarly, there are several factors to consider when selecting... week planning an experiment, even a relatively simple experiment. However, by taking this time, you can...result? Is there a tractable question or are you simply describing a phenomenon? Exploration can be important...Plus, presentations benefit from practice. If you simply recycle slides from the last time you gave a presentation...

...Fluorescent proteins (FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate...fluorescence when illuminated with blue light (1). Similar to PA-GFP, scientists later developed red fluorescent...photoactivatable fluorescent protein for your experiment Similar to performing experiments with standard FPs, before...live imaging of proteins, organelles, and cells. Similar to FPs, PA-FPs allow for non-invasive labelling...

...using the csRNP method is as simple as doing routine PCR assays. Here is a simple to-do-list to follow when...spectrometry to identify the interacting proteins. Similarly, in ChIP-seq assays, chromatin fragments that ...successfully knock-in non-expressed genes with similar efficiency, Dewari et al. 2018). Using the first...

.... To start the AAV packaged on request process, simply navigate to your plasmid of interest. If it’s eligible...cargo you requested at the quality you expect. Similar to our on-demand viral service, we expect titers...There are no quotes or special processes involved; simply request your plasmid of interest in your desired...

... used as the first letter of a plasmid name and simply denotes that the object is a ‘plasmid’. “p” is ... empty backbone name in your plasmid name. This simple piece of information can often convey many important...pBACKBONE-Flag-hGene-EGFP or pBACKBONE-Flag-hGene(wt)-EGFP These simple rules allow any scientist to know what is in a ...

...edited strand. The first iteration of prime editor simply fused the wild-type Moloney Murine Leukemia Virus...or exceed PEmax editing efficiencies for short, simple edits. PE6c and PE6d further evolved the Tf1 and...pegRNAs, but combining PE7 with epegRNAs resulted in similar or reduced editing efficiencies, likely because...Ravisankar, P., Ward, C. C., Levesque, S., Jing, Y., Simpson, D., Zhao, A., Li, H., Yan, W., Goudy, L., Schmidt...

...and cell lines. Although ChIP-seq is a relatively simple and robust protocol it does have limitations. The...change offers a significant improvement and simplification of the original technique, specifically in ...CAPTURE Additional Resources on Addgene.org Find simple lab protocols Browse CRISPR plasmids for isolating...

... DA in live animals (Patriarchi et al., 2018). Similar to calcium and glutamate sensors, the new dLight1...s lab has created the GRABDA sensor based on a similar strategy: a GFP variant bound to a DA receptor,...cyclase in caudate nucleus of rat brain, and its similarity to the “dopamine receptor”." Proceedings of the...

...Drawings is similar to using PowerPoint to draw figures. It’s part of Google Drive so it has a similar interface...figures quickly. Inkscape Inkscape is the most similar to Adobe Illustrator out of the three options for...

...control over neuronal activity. While they serve similar purposes, both techniques offer researchers different...resolution as receptors can be turned on and off simply with the presence or absence of the correct wavelength...flexible as it relies on designer ligands that can be simply introduced by injection. The main challenge in ...

...Interspaced Short Palindromic Repeats. That contrast of simplicity and complexity is reflected in the biology, too...into a single strand that folds back on itself, simplifying the CRISPR-Cas9 system for laboratory applications...post on the structural biology of Cas9. Want a simpler guide to keep at your desk? We’ve got a CRISPR ...

... the Cellosaurus STR similarity search tool (CLASTR) to find cells lines similar to your STR profiles ...CLASTR results by minimal number of markers and similarity score match. This tool’s algorithm can be adjusted...

...Engineering the Plant Genome Using CRISPR/Cas9 pSiM24: Simplifying Plant Genetic Engineering Resources on Addgene.org...Addgene.org Find Plant Vectors at Addgene Find Simple Protocols for Introductory Lab Work MoClo Plants...

...provides a combinatorial indexing strategy that simultaneously tracks clonal history and cell identity at ...that cells with common ancestors tended to have similar reprogramming outcomes. For example, cells with...with shared CellTagD13 tags often had similar gene expression pattern changes and reprogramming outcomes...

...equipment. Reporters for studying gene expression have similar drawbacks, often requiring the destruction of plants... events and am currently planning to use it to simplify my work with an Arabidopsis mutant that can’t ...unfamiliar with these mutant plants. Using RUBY to simplify the identification of these mutants will speed...

...resistant to vaccine serum responses. This result was similar to studies that used live virus (Cele et al., 2021... shocked [with these results] because we were simulating the live virus experiment,” says Balazs. “But...neutralization rates of B.1.351 pseudovirus was similar to that of pseudoviruses representing a bat-derived...

...ve ever wondered how they are different from or similar to the other Ts, we have your answers. If you know...into a cell by a viral agent. Transductions are similar to transfections, processes which introduce foreign... Transfection and transduction seem to achieve similar outcomes, so why and when should you opt to make...

...multifunctional fibers, embedded, and validated simultaneously in the brain and the intestines of mice. The...organisms, including the Anikeeva lab’s work on a similar neural fiber (Park et al., 2019). This provided... To operate both types of implanted fibers simultaneously, a mini-Bluetooth system named NeuroStack was...

...Viruses are simple: a genome packaged in a protein shell (Taylor, 2014). They’re so simple that we can... even decide if they’re alive or not. Yet these simple, small particles have quite the outsized impact...

... multiple sgRNA constructs to be introduced simultaneously in a single plasmid. The functionality of the...of S. cerevisiae and E. coli. The utility of simultaneous and multidirectional multiplex control via scRNA...Multiple scaffold RNAs can be designed to allow simultaneous, multidirectional and tunable control of gene...

...the Gal4/UAS system. This incredibly useful, yet simple system allows you to systematically study gene ...downstream of a tissue-specific promoter sounds simpler, this modular system saves you from having to create...compartment, you could express them individually or simultaneously. Do note for control purposes that there are...

... about operating the rest of the car as well. Similarly, cell free expression systems contain all the ...students to use in the classroom because the students simply need to add water to express a variety of proteins...Addgene.org Browse our protocol videos to learn simple lab techniques Check out our molecular biology ...

...levels for 6 days - after just 15 minutes of FUS.  Similar constructs to the one above can be made for expressing...But why might you need a second GFP antibody?  Similarities:  Generated against full length GFP and converted...the group rationally designed GFP variants with similar capabilities. If you’re looking for a super stable...

...disappears from the video, and prepares a coffee, or simply moves away from the desk to switch off a few minutes...GA, O’Reilly CM, Lofton ME, Catalán N (2018) Ten simple rules for collaboratively writing a multi-authored...journal.pcbi.1006508 Vicens Q, Bourne PE (2007) Ten Simple Rules for a Successful Collaboration. PLoS Comput...

...amplification protocol, but the overall workflow is similar for most libraries. Here are the key points you... For libraries where the plasmids are all very similar sizes, like a gRNA library, this is less of a concern... hope you’ll agree with me that they look very similar to each other - that’s exactly what we want! At...

...TCSs have been identified in bacteria, and with similar optimization methods, many of these systems could...see light. Levskaya A, Chevalier AA, Tabor JJ, Simpson ZB, Lavery LA, Levy M, Davidson EA, Scouras A, ...genetic edge detection program. Tabor JJ, Salis HM, Simpson ZB, Chevalier AA, Levskaya A, Marcotte EM, Voigt...

...proteins they identified have little sequence similarity to previously identified Cas13a-c orthologs, ...mammalian system. Since Cas13d is functionally similar to previously discovered Cas13 enzymes - what makes...their cleavage pattern: EsCas13d produced very similar cleavage products even when guides were tiled across...

...promoter and ceases repression of the target gene. A simplified lac inducible system is found in many bacterial...between cell types or organisms, promoters are similarly variable. Bacterial promoters only work in prokaryotic...related species from which they were derived. Similarly, the various eukaryotic cell types (mammalian,...

...create a genome-wide library capable of performing similarly to Cas9 screens in a compacted library. Overall...genes within a single cassette provides increased simplicity for combinatorial genetic screens, and this optimized...Penn State Protein Ladder  by: Rachel Leeson This simple, inexpensive plasmid-based ladder system is a cost-effective...

...Identifying the Issue You can’t fix an experiment by simply running it unchanged over and over again, and the...was not trying even harder to start earlier, but simply moving all my prep work to the day before, which...remember what's been added and what hasn't. Developing simple habits like these can consistently reduce errors...

...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 ... this is an Antibody 101 post, so we'll keep it simple. If you follow the two basic guidelines below, ...

...facilitates the reproduction of the structure in a similar way to the biological protocol, and assembly designs... young movement of sharing open hardware has a similar goal. The sharing of modules, ideas, and blueprints...Nibha Akireddy, Chew Chai, Manu Prakash, "Squid: Simplifying Quantitative Imaging Platform Development and...

...suppressor screens These types of screens are very similar to forward and reverse screens, except that they... which are worth considering. RNAi screens are similar to CRISPR, but instead of knocking out the genes...temperature to exacerbate a mutant phenotype, very similar to enhancer/suppressor screens. Selections Selections...

...system work? Mendenhall: The system is really quite simple but it offers improvements in efficiency and throughput...right thing to do to put these out there. It’s a simple solution. It would be silly not to share it as ...on many other labs that may be doing something similar. There may be other variants of this that are better...

...together, thus repairing the break. This is a simplified, streamlined version of this pathway and does...regions of hundreds of base pairs or more can simultaneously introduce a pair of chromosome breaks, and ...deletions) if NHEJ joins the distal ends together. Similarly, it may be possible to direct insertion of an ...

...threonine kinase with a similar 3D structure to MEK1, but Raf1 and CDK5 have less similarity. For these three...versions of MEK1, MEK2, Raf1, and CDK5 all behave similarly to their endogenous counterparts. Additionally...

...absorb the excitation light less efficiently. Similarly, decreasing the power of the excitation light ...results in a loss of fluorescence, which, in its simplest form, can be described by a mono-exponential decay...intensity decay of a fluorophore does not follow a simple mono-exponential decay (Shaner et al, 2008; Bindels...