...V-K CRISPR-transposon systems function in a very similar manner to Type I-F, but instead employ a Cas12k...in single-plasmid INTEGRATE (pSPIN), which offer simple transformations for smaller cargos, and the ability... with the V. cholera INTEGRATE system follows a simple workflow: Clone spacer(s) into the CRISPR array...sequencing While CRISPR-transposons are powerful and simple to use, users should keep aware of a few important...

...the plant genome using CRISPR/Cas9 Using pSiM24 for simplifying plant genetic engineering Using arabidopsis...seed growth on antibiotic-containing media) A similar process occurs in nature; Agrobacterium tDNA insertion... sweet potatoes (Kyndt et al., 2015)! It sounds simple, and it pretty much is, but there are some tips...

...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...

...hemagglutinin (HA), myelocytomatosis viral oncogene (myc), simian virus 5-derived epitope (V5), the synthetic peptide...ability to distinguish weakly expressed and/or similar proteins Through their multiple, highly specific...standard tags and can be used to distinguish highly similar proteins. For example, N-cadherin (cadherin-2) ...belongs to a huge family of proteins that are highly similar in terms of sequence and thus very difficult to...

...mammalian plasmids, mammalian vectors are even less similar to bacterial ones. The mammalian analogue of bacterial...stable mammalian cell lines, but it isn’t common for simple transfection experiments. Reporters Finally, in... between. For example, if you are using E. coli simply for easy cloning and quick DNA production, you ...have taken up an antibiotic resistance gene they simply can’t turn on! Wait, what even counts as a shuttle...

...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...

... 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...

...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...

... 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...

...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...

...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...

...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...