...itself. Among other things, science communication comprises broadcasting, science writing, and certain aspects... get on stage for a Science Slam – a sort of competitive TED talk – to win the audience’s applause with...photography and participate in a science photo competition. Or nominate yourself as an educator for a short...

...structure of the hairpin-DNA complex is thought to destabilize the transcription complex, initiating cleavage ...mechanism, whereas Polymerase II termination is more complex and involves two RNA polymerase-associated proteins...

...who are usually antagonistic and impolite, Complainers, who spend most of their time moaning about their...with negative comments (hostile-aggressives, complainers, know-it all experts, wet-blanket negativists...support from your colleagues or other supervisors to complete your work. Whether your supervisor is a negative...

...white dot growing on the solid medium. This dot is composed of millions of genetically identical bacteria ...

... fluorescent signal throughout the reaction is compared to a standard curve of amplified DNA of a known...The cycle in which the unknown DNA is detected compared to the standard curve can be used to determine... to note however, that multiplex PCR is more complicated to develop and is often less sensitive than PCRs...

...proteins on the cell surface. Figure 1: Comparison of common antibody, HCab, and nanobody. For more...starting library size (up to 1012 members) in comparison to other display systems using phage and yeast...the end of the mRNA molecule, forming a ribosome complex consisting of the translated protein (sybody), ...

... strategy uses new combinations of PE protein components (Figure 1), an altered epegRNA delivered with...protein-coupled receptors (GPCRs) are critical components of many physiological pathways and represent ...et al., 2024). Each plasmid expresses all the components of the biosensor in optimal proportions from ...

...screens have gotten larger, easier, and more comprehensive thanks to the consistent development and improvement...vectors is how the cargo — in this case, the CRISPR components — are expressed. Unlike lentivirus, cargo delivered... away from the comfort of Cas9, and opt for a completely different Cas effector, like Cas12a (Cpf1). Even...

...RNA (pegRNA). The pegRNA consists of a target-complementary spacer, a scaffold sequence, and an extension...accounting for some of the inherent inaccuracies of computational prediction models. In addition, we provide options...Switzerland), where he developed a strong passion for computational biology. His expertise includes applying machine...

...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...the material used for inoculation. To further complicate matters, plasmid copy numbers are often ambiguous...

...also is also called Bimolecular Fluorescence Complementation (BiFC). 8 points to keep in mind when choosing...divided by 1000. In many cases the brightness is compared to that of EGFP which is set as 1. Some proteins... BD bioscience. ilovegfp – a site with very comprehensive data sheets on many FP variants Plasmids 101...

...engineered DNA-binding molecule such as the CRISPR complex is expressed in cells to be analyzed, the Fujii...another "in vitro" form. In this form, the CRISPR complex, consisting of a recombinant dCas9 protein and ... a genomic locus through a specific gRNA-dCas9 complex. When combined with next-generation sequencing,...

...No examination of the state of science would be complete without the inclusion of some interesting case...Prashers will find it increasingly difficult to compete (for funding, etc.) with that produced by the confident...the U.S. environment, and a decline in the competitiveness of U.S. science and technology.” This last ...

...your samples, the relationships between cellular components, and the accessibility of your target epitope...work by dehydrating and precipitating cellular components. This method of fixation is good for epitopes...away your target (for example, some cytoskeletal components) and you want to fix and permeabilize in one ...

...cells can decrease the number of fluorophores, compromising the photon budget. Likewise, the switch from...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...

...importance of the entire GFP protein, which is almost completely required to maintain fluorescent activity; very...may not apply. Table 1: The specific mutations comprising common fluorophores Fluorescent Protein Mutations...chromophore to form with an indole rather than phenol component (cyan derivatives) Y66H Blue-shifts the wavelength...

...laser cleavage and an RNA aptamer that bound the compound malachite green (MG) to create a non-protein based... 4-hydroxybenzylidene imidazolinone (HBI)-like compound allowing it to fluoresce, the Jaffrey lab asked...production have also made aptamers an important component of high throughput assays with microarrays and...

...decreased p-values of known ricin regulation genes compared to results generated using non-targeting guides...full-length (19-20 bp) or short (17-18 bp) guides. When comparing the toxicity of guides that have 1-bp mismatch...off-target sites, short guides had reduced toxicity compared to full-length guides. Reduced toxicity of short...

...1997). We have a lot of plasmids that contain components of the SB transposon system including those from...but think again. Spaghetti squash is actually a component of the fly actomyosin cytoskeleton that regulates...regulates cellular contraction. This network is composed of actin and myosin which interact with each other...

...C) installation by base editors. Pathways that compete with the desired outcome are not pictured. ...Plant base editor platforms and their molecular components. Adenine base editors for plant We developed...editors. Plant CBE and ABE with expanded PAM compatibility All of the base editors mentioned above are...

...are differentially inactivated by nonspecific complement mechanisms in serum. Sometimes this is due to...sugars end up in the envelope and are targets for complement-based killing by anti-alphaGal antibodies. The... as described above and is inactivated by the complement system in serum (DePolo et al., 2000). Other ...

...the anticodon,  a three nucleotide sequence complementary to the mRNA sequence. The 3’ end of  the tRNA...resources (Athey et al., 2017) and many DNA synthesis companies, such as IDT, Genewiz and GenScript, offer tools...the other while other codons are under expressed compared to others. Image from Hiss et al., 2017. Most...

... amplify well and others don’t. The size and composition of your library can drastically impact what conditions...suffice…and the answer is no. Due to the size and complexity of pooled libraries, NGS is the best way to validate... plasmids didn’t amplify after your screen is completed!  Producing Your Lentiviral Library Once you've...

...both Addgene’s founding and our growth into the company we are today. As part of our 20th anniversary celebration...pages allowed researchers to easily access and compare their options for tools such as empty backbones...Addgene YouTube channel to add a strong visual component to our educational resources. From 2013–2023, ...

...ordering the sgRNA is simple and easy. Several companies supply gRNA sequences, such as GenScript, ThermoFisher...order them yourself through whatever DNA supply company you currently use. You can also find many previously...explored and it allowed control over each individual component of the system. However, it is easier to directly...

...line identity and purity making it easier for comparable studies to be performed on a cell line even decades...passage number, cell line, and date, which you complete each time you passage the cells (as well as recording...through working for a variety of pharmaceutical companies including GSK, AstraZeneca-MedImmune, and Infors...

...needs a standard against which the results can be compared; results from an experiment lacking the proper...baseline/standard against which other samples can be compared. The Empty Vector Control (mentioned above) could... where expression of that gene is knocked out completely. These controls may or may not be plasmid-based...

...promoter present in your plasmid backbone must to be compatible with the type of RNA that needs to be made: if...speaking, promoters in bacteria are less diverse and complex, having fewer parts than those in eukaryotic cells... repression in the presence of glucose or by competitive binding of the anti-inducer fucose Weaker. Commonly...

... in other lanes contaminating your sample. To accomplish this, it is best to skip lanes between samples...

...trace file (.ab1 file) provided by your sequencing company or core facility The corresponding primer sequence...

...reagents is important to ensure the accurate composition of your solutions and buffers. Now that you have...

...then following transduction, the partial genomes complement each other to restore the full-expression cassette... has equal or higher transgene expression when compared to using a single over-sized vector, but not always... one of the transgene’s endogenous introns or comprised of non-endogenous sequences, such as a 77bp long...

...recombination intermediates can then resolve to complete the DNA repair process Homology directed repair...is elongated and ligated to fill the gap, thus completing SDSA.   Break-induced repair (BIR) pathway BIR... uses reverse transcriptase to generate the complementary ssDNA. Easi-CRISPR was shown to increasing editing...

...get people to remember you). What better way to accomplish all of these things naturally than to give a ...simplicity of their slides and how the slides complement the spoken message. Learn from their level of... topics that are unfamiliar to you because the company you are visiting works on them. Of course, if some...

...regularly. Dystrophin and genome editing Since complex oligonucleotide treatment comes with many challenges...desirable. Should stem cell editing occur, the CRISPR components would only need to be expressed for a short period... with vastly reduced inflammation and necrosis compared to unedited animals. In grip strength, specific...

...system, Sanes and Lichtman also developed a complementary Flpbow construct that is functionally similar...technique will provide important insights into the complicated physical organization of the brain and other ...Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells...

...that you have is the expected size, or that it is composed of a backbone and insert of expected sizes. This...

...positions he has held in his career, his favorite company tradition, and Addgene's role in the scientific...

...Gene knockout with CRISPR technology is usually accomplished by Cas9-mediated dsDNA breaks: following a cut... accessibility due to chromatin status). No computational prediction is ever perfect, but this can decrease...to conduct genetic screens, however, a library composed of the latter would likely be more effective, ...

...stimulation. GPCR signaling GPCR signaling is a complicated process involving many downstream signaling mechanisms...applying optogenetic tools derived from direct comparative analysis of microbial opsins. Nature Methods,...