We narrowed to 933 results for: COMP
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TypeBlog Post...experts that complement your (scientific) skill set and is therefore very useful for complex projects. It...collaborations are becoming increasingly important as more complex approaches to scientific questions become popular...especially when considering small to mid-sized companies and organizations. Addgene as a nonprofit organization... worked with people outside the department to accomplish a variety of goals. For example, I worked with...provided a crash course in this discipline. Getting companies to come to these types of events can be very tough...First I needed to find the right contacts at the companies and then convince the contacts to give me some...repetition. I learned to look at the event from the company’s perspective, focus on the message and to keep ...
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Plasmids 101: Shuttle Vectors
TypeBlog Post...function in unrelated host species, even from completely different kingdoms of life. That’s convenient... for researchers who want to do experiments in complex eukaryotic cells, but would prefer to grow, clone...replication begins, so a bacterial plasmid needs a compatible ori that will be recognized by its host’s replication...into plant genomes. Eukaryotes are a bit more complicated. Yeast vectors may contain an autonomously replicating...increase cellular uptake, but this approach requires compatible cell lines that express viral machinery. Alternatively...may be sufficient for your needs even without a compatible replication site. Regardless of the species you...vector to persist in... You need... Bacteria a compatible ori Yeast an autonomously replicating sequence... -
Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs
TypeBlog Post...with fluorescent proteins (FPs) localized to compartments other than the cytoplasm. That is, unfortunately...positioning of the FP sequence relative to cell compartment targeting sequences, which often have absolute...that FPs are often poorly suited for cellular compartments other than the cytoplasm. Generally, FPs evolved...subcellular environments, including the organelles that compose the secretory pathway, endocytic vesicles, mitochondria.... Many of the resident proteins within these compartments undergo significant post-translational modifications...inter-chain disulfide bonds between FPs blocks Golgi complex-localized fusions from properly exiting the ER ...and spectral characteristics of the moxFPs are comparable to the non-optimized parental proteins. Our recent... -
Understanding GPCR Signaling Using TRUPATH
TypeBlog Post...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...They further used TRUPATH to analyze a panel of compounds across the transducerome on a single GPCR, showing... -
The Fluorescent Vegetables in Aptamer Soup
TypeBlog Post...locations using Cas9 and track the localization of the complex. Broccoli, the other green fluorescent aptamer,...investigate RNA assembly. The split aptamer is composed of a “Top” and “Bottom” portion, each fused to... RNA. Fluorescence is observed when these RNAs complex and the entire aptamer can hybridize and fold. ...uniqueness of the photostability of the Corn-DFHO complex. For dessert (last but not least), the only fruit-named...Other groups have used Mango to study RNA-protein complexes (Panchapakesan et al., 2017) and short non-coding...identified through microfluidic-assisted in vitro compartmentalisation for their high binding affinity to TO1-biotin...of these variants exhibit increased brightness compared to Mango, and all are photostable and useful for... -
An Integrin Antibody Toolkit from IPI
TypeBlog Post...conformational complexity There are 24 known mammalian integrins, all heterodimers composed of one of 18...are essential cell surface receptors involved in complex signaling pathways and the linkage of intra- and...-integrin toolset IPI’s anti-integrin toolset comprises 26 recombinant antibodies (rAbs) either produced...-mimetic sequences that imitate this motif and compete with both small molecule integrin inhibitors and...their targeted integrin subtype, even when in competition with other similar integrins, as reported by ...demonstrated that anti-integrin αVβ5.9 “essentially completely blocks all adhesion,” inhibiting the function... rAbs. The other eight are chimeric antibodies composed of rat or mouse variable regions fused with rabbit... -
Evolution of Lab Techniques
TypeBlog Post...when. Compiling the identity and location of a given nucleotide, a researcher (or a simple computer program...in Neuroscience at Harvard University. Just as computers, cell phones, and cars become more technologically... fluorescently labeled probe. If the DNA is complementary to the probe (for example, has the sequence ...be quantified by the strength of the signal in comparison to a control sample. Southern blotting applications...in standard PCR. This allows the researcher to compare different DNA sequences to see which is more abundant...mapping to investigate DNA sequence content by comparing the fragments created by different restriction... sequencing of many samples at once for later comparison. These next-generation sequencing techniques ... -
FPbase: A new community-editable fluorescent protein database
TypeBlog Post...non-trivial, however, as FPs are tremendously complicated entities with a large range of characteristics...about FPs than is currently represented at fpvis (complete spectral information, accession numbers, sequences... (PostgreSQL), that aims to eventually be a comprehensive community resource for information about fluorescent...Protein Property Visualization allows for visual comparison of FPs and how they relate to one another. . ...FPbase Fluorescence Spectra Viewer can be used to compare excitation and emission spectra when available,...you or your lab use frequently) for the sake of comparison or ease of retrieval. These collection pages...database relating to that set of proteins. For computer-savvy users looking to incorporate FPbase data... -
Plasmids 101: NGS Quality Control for Pooled Libraries
TypeBlog Post...transformation First, use enough DNA and competent bacteria to ensure all components of the library are represented...represented. Typically, Addgene uses electrocompetent Stbl4 cells when performing amplifications, and we use electroporation...may be amplified preferentially, skewing the composition of the final recovered DNA. This bias can be ...need to count how many times each of the library components is found in the read list. If you also sequenced...sequenced the pre-amplification DNA, you’ll compare the numbers before and after. However, because there are...give you a bell curve of the abundance of each component of your library. You should look for two things...making sure few/no plasmids have dropped out, and comparing pre- and post-amplification sequencing results... -
Synthetic Photobiology: Optogenetics for E. coli
TypeBlog Post...consistent, and controllable pulses of light. Complicated patterns of light intensity and wavelength can...not E. coli. They fall under the heading of two-component systems (TCSs). TCSs consist of a histidine kinase...created the first E. coli light-sensitive two-component system (TCS). Levskaya et al. fused a photosensory... image, as well as a genetic method for the computational problem of edge detection. Tabor et al. subsequently...gene expression. An E. coli light-sensitive two-component system (TCS). A hybrid histidine kinase/photosensor...experimented with the promoter strength of various components, removing inducible promoters that could crosstalk...expression levels are optimal. These tools are compatible with many strains of E. coli, as well as with... -
Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility
TypeBlog Post...larger datasets and use computer based analysis techniques, an important component of reproducibility is... for preclinical studies, as reported by drug companies attempting to apply the findings from these studies...research. I’ve talked to many researchers who have complaints about the unreliability of antibodies. IPI is...technologies to your own research needs. Other companies like NEB and ATCC can provide you with additional...sifting through paper lab notebooks). Once you’ve completed your experiment, you’ll need to analyze your results...can understand your analyses and repeat them. Companies like Code Ocean allow researchers to share code...submit designs and transparently reveal them upon completing an experiment. There’s not necessarily in-principle... -
Sharing is Caring: But How to Distribute Open Hardware?
TypeBlog Post...achievement, the complexity of the device increases, as does the price. With this increased complexity comes the...consumer-grade components. In this field, often referred to as 'frugal science', existing components are adapted...are constantly being developed which combine computational processing with biological protocols and revolutionary... rarely provide the construction plans or CAD (computer aided design) drawings of their devices, which...around the use of commercially available tools and components. Rapid prototyping techniques such as 3D printing...and can allow for innovative organizations and companies to provide problem-solving platforms. ... -
3 Challenges in Plant Synthetic Biology
TypeBlog Post...previous blog post about how they started their company here. The first transgenic plant was engineered...cannot build what we do not understand, and the complicated and unique metabolic processes of plants are ... Our color changing flower involves two basic components: the pigments (anthocyanins), and the plant circadian...also identified and characterized mechanisms and components of the circadian rhythm in petunias. Because ...before us, it’s possible for us to combine these components to develop flowers that change color. Without...Without this basic research – this comprehensive understanding of the genetic pieces underpinning these mechanisms...plant synthetic biology. There are already two comprehensive plant cloning kits available in the repository... -
Antibodies 101: Selecting the Right Antibody
TypeBlog Post... as these immune complexes are smaller than a primary:secondaries:conjugates complex. If you are trying...and therefore each target protein. This can be accomplished by using a panel of primary antibodies conjugated...tissues and cell lines; orthogonal strategies that compare antibody assay results with those of antibody-independent...transcriptomics; independent antibody strategies that compare the staining patterns of multiple antibodies against...that express a tagged version of the antigen and compare staining of the antibody to that of the anti-tag... strategies that isolate an antibody’s immune complexes and confirm interacting partners by mass spectrometry... guaranteed to work. Antibody-based assays are complex and antibodies may perform differently in different... -
Live and Let Dye: Self-Labeling Protein Tags
TypeBlog Post... chemistry. Although these multi-component systems can be complicated to implement, they can be highly...Various Few Complicated Non-natural amino acids 1 Various Many Complicated Applications...STED) nanoscopy. Scale bar, 2 μm. Inset shows comparison with standard confocal microscopy. Image adapted... assembly-regulating proteins in vitro reveals complex events. a) Micrographs of a fluorescently-labeled...159 aa) are smaller than HaloTag (295 aa) – for comparison, common fluorescent proteins are typically 230...Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and SNAP-tags. Cell Chemical...Ulrichs, H., Gaska, I., & Shekhar, S. (2023). Multicomponent regulation of actin barbed end assembly by ... -
Educational Planarian Tools for Teaching Developmental Biology Techniques
TypeBlog Post...worm amputations. They also created an online companion resource called “cutting class” which provides...Digoxigenin-labelled Uracil) that will bind to complementary mRNA within an embryo or tissue. Once the riboprobes...specific antibodies that detect the mRNA-riboprobe complex are added to the tissue. The last step involves...cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue. Students...organism of interest, use the RNA to create a complementary DNA (cDNA), and then use PCR and molecular cloning...performing these experiments have a good baseline for comparison. Performing WISH using these plasmids can result...one strand of the siRNA and are directed by complementarity to the remaining single stranded siRNA to bind... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog Post...RNA is a structural and functional component of the ribosome complex and Dr. Steitz quipped that RNA in...development zone so many science and technology companies and start-ups choose to locate there. I made time...was the outdoor market where one can shop for components of traditional Chinese medicine. There were hundreds...Lowe lab have allowed them to begin understanding complex regulation of and by tRNAs and tRNA-derived RNAs...replication with no proteins). The chemistry is complex and this work requires an interdisciplinary team... they do it. Of course, No conference would be complete without CRISPR Michael Terns from of the University...in the ribosome is in a complicated conformation “not like DNA which has a boring structure.” That certainly... -
3D Printing Meets CRISPR Cas9
TypeBlog Post...realized how perfectly flat the complex is on this surface”….where the complex is docked on the membrane of...using 3D printing technology. The tagline for our company is “…where molecules become real”. We sometimes... the real thing. Nevertheless, the models are compelling, and I encourage you to hold one. Why do we ... They then start talking to themselves as they compare the features displayed on the model with their ... close to its reported cleavage site on the complementary DNA strand. And this is why we love working ... that was not “poised for cleavage”. Jacob computationally added the non-template strand to the Jinek ...double-stranded DNA, recognize a PAM sequence, allow the complementary base pairing between the guide RNA and the target... -
Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria
TypeBlog Post...when studying them since their complete knockdown is lethal. Incomplete or titratable knockdown, however...generations. Figure 1: Mobile-CRISPRi's modular components. Image from Peters et al., 2019 with permission... Top: A Tn7 transposon containing the CRISPRi components and a plasmid with the Tn7 transposition genes...ability to synthesize diaminopimelic acid (DAP), a component of the cell wall. Once in the recipient cell, ...Bottom: An ICE element containing the CRISPRi components is transferred to recipient bacteria by bi-parental...was variable. They measured transfer rates by comparing the number of colonies that grew on a selective... gRNA were more sensitive to trimethoprim when compared to control gRNA treated or untreated bacteria.... -
Plasmids 101: 5 factors to help you choose the right cloning method
TypeBlog Post... experiments are also generally comparable so the best way to compare costs is to look at the prices of...In today’s post, we will focus on a high-level comparison of popular cloning methodologies, with a special...least 50 cloning reactions. The ligase cost is comparable to that of the restriction enzymes and overall... in a price of a few dollars per reaction. By comparison, Gibson assembly can cost over $15.00 per reaction...more streamlined, because they will come with comprehensive and tested protocols, as well as premixed and..., like Gibson or SLIC, since one can pool the components and define their final order through sequence...Golden Gate assembly allows one to simultaneously complete an ordered set of scarless ligations following...