We narrowed to 1,075 results for: ica
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TypeBlog Post...repurpose popular GFP reporter lines for more sophisticated experimental manipulations, saving the time ..., 98% of TdTomato expressing cells were GFP+, indicating a robust, but specific system. T-DDOGs also successfully... types of proteins. They started with another biological workhorse, Cre recombinase, which induces recombination...systems are easily adaptable to neuroscience applications, including optogenetics, and they should make...Tang note that their scaffold system is likely applicable to many types of proteins. For proteins with ...Could Cas9-DOG be developed soon to allow genome modification only in a small subset of cells? Other proteins... PMCID: PMC4096992. Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “...
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Plasmids 101: Golden Gate Cloning
TypeBlog Post... work around this is to "domesticate" your fragment: PCR-based amplification can be used to create silent...and exciting applications, this system is an incredibly powerful tool for cloning complicated constructs...plasmid (Option 1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning...restriction sites that may not be amenable to "domestication", you might want to consider using an alternative...design of flanking overhangs. Although there are theoretically 256 distinct flanking sequences, sequences that...high-efficiency step. References: Sequential amplification of cloned DNA as tandem multimers using class-IIS... -
Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them
TypeBlog Post...generated in vitro using synthetic genes that are typically expressed from a plasmid or from an integrated...cells, for antibody production and subsequent purification. Academic laboratories and companies have already...share, making recombinant antibodies the most practical choice. Hybridomas are much tougher to ship, often... often lack sequence data, and may genetically drift over time. Antibodies made from animals are limited...the ease with which scientists can now improve genetically-encoded research tools, it is shocking that the...Begley CG, Ellis LM (2012) Raise standards for preclinical cancer research. Nature 483:531–533 . https:/...Edition. 2104. Cold Spring Harbor Laboratory Publications, New York. Helma J, Cardoso MC, Muyldermans ... -
Corralling and Cornering the Goldilocks Antibody
TypeBlog Post...the best, or even which one will work for your application? Like many scientists, I have spent hours sifting...researchers to identify and isolate very specific biological targets. However, as popular and useful antibodies...produced from a new set of animals), and wrong application (different experiments/conditions can change ...differently over time), compatible with the desired applications (such as immunohistochemistry (IHC) or western...colleagues! Dr. Wu and his lab members (like me!) are dedicated to creating a public and accessible database for...antibodies, creating a potentially unlimited supply of identical antibodies (See Fig 1.). rAbs are a relatively...a research assistant in lab of Zhuhao Wu at the Icahn School of Medicine at Mount Sinai. References ... -
A Guide to Designing a Scientific Poster: Content, Layout, and More
TypeBlog Post...very useful in the event that you will not be physically present at your poster at all times, so other... sections should be as concise as possible, communicating only the most important results to your audience...have the advantage of being able to verbally communicate your work, so you don’t need every little detail...readers can navigate your poster and follow the logical progression of your work, and you can accomplish...offers free poster templates, although most are vertical. Large central box. Another commonly used and...presentation. Portrait. This style displays a poster vertically rather than horizontally. Instead of displaying.... Shapes. In this type of poster, written and graphical information are inserted into larger circles or... -
Pathways Over Time Plasmids Engage Students in Functional Genomics Research
TypeBlog Post...undergraduate students (1, 2), but educators face significant challenges in designing suitable projects. In... become very different due to a whole genome duplication and extensive rearrangements in the S. cerevisiae...overexpression vector. This plasmid contains origins of replication and selectable markers that allow students to...functional MET genes to grow.) Students then use replica plating to determine if expression of MET/Met genes...met deficiencies (manuscript in preparation), indicating that Met protein functions have been largely ... Students who participate in the course show significant gains in both content knowledge and research ... research into undergraduate courses and in communicating science to the public. You can find out more... -
Plasmids 101: Modular Cloning
TypeBlog Post...production and subsequent necessary modification of these chemical products. The flexible nature of MoClo... Gate cloning is MoClo. That’s because MoClo specifically refers to the organizational schema described...organization schema MoClo can be broken down into three hierarchical levels: individual DNA parts, transcriptional...together. They can express one or more gene(s), and typically begin with a promoter and end with a terminator...multi-purpose it can be. Let’s review a list of MoClo applications. Genetic circuits Most things in biology are...microbially or in cell-free systems. These pathways typically involve multiple steps in which a precursor molecule...genotype and phenotype is best described with “it’s complicated.” Applying MoClo Interested in using MoClo? You... -
5 Science Rap Videos & Their Creator: Tom McFadden
TypeBlog Post...Lacks - the African American woman whose cervical cancer cells became a mainstain of biological study without...advice on how to dive into new forms of science communication. Click here to listen to the interview with ...fossil rocks and their importance in understanding biological history. 4. Please Don't Kill My Hive Beautifully...implement the next generation science standards and communicate science. Learn more about Tom on his youtube ... -
Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity
TypeBlog Post... more than 10 modules often rely on two-step hierarchical approaches using different Type IIS restriction...stability, and buffer compatibility have placed practical limits on single- or two-step assemblies. We have...upon growth on LB/Cam/X-gal/IPTG agar plates, indicating successful reconstruction of the coding sequence... While T4 DNA Ligase, the mainstay of most biotechnological cloning efforts for over 50 years, prefers...-Crick base pair substrates, it demonstrates significant activity on some mismatch-containing pairings... sequence-dependent ligation efficiency and identification of mismatch-prone pairings. Using these 4 bp...Kucera is a Principal Development Scientist of Applications & Product Development focusing on DNA cloning... -
Of Myc and Men
TypeBlog Post...recognized by its antibody without additional chemical modification steps and when the epitope was within the...antibodies for every potential new target and basically impossible if you wanted to distinguish between...C-terminus post-translationally, which turned out to be critical for antibody recognition. As such, the peptide...interest and proteins tagged with it needed to be chemically treated before the antibody could detect them... the N-terminal half of the antigen differs significantly between human and the other species, suggesting... flexible - it could now be used in way more applications. Fig. 1 Making a Protein Tag (Abridged... of new versions of Myc1-9E10 mentioned above basically involve taking the Myc1-9E10 variable regions ... -
The Scientific Conference Poster Session: Tips for Success
TypeBlog Post...example, if you’re a virologist presenting at the American Association for Cancer Research Annual Meeting...therapeutic strategies to target it, than you typically would at a microbiology meeting. Create an outline...lab mates, and PI can provide feedback on your technical presentation, but those outside of your field ...don’t fully understand the question, ask for clarification. Having them reframe or elaborate their thought... through the list at the session so you can strategically use your time. Don’t have access to a list of...titles to find the ones you’d like to revisit. Strategically traverse the sea of posters If you come prepared...system, and is interested in a career in science communication. Resources on the Addgene blog Read other... -
Addgene's Cancer Collection Pages Connect the Research Community
TypeBlog Post... the pharmaceutical industry for years. They have proven very difficult to target therapeutically due ...al. "Systematic identification of signaling pathways with potential to confer anticancer drug resistance... millions of people die from cancer-related complications each year. Despite these grim statistics, new... cancers, and synthesizing compounds to bind specifically to their mutated residues. For Ras researchers...any of these genes through several different applications such as fluorescent protein fusion studies and...represented, making the results of any such study as clinically relevant as possible. Connecting the research... -
PITChing MMEJ as an Alternative Route for Gene Editing
TypeBlog Post...another method for making targeted modifications. A recent publication by Zhang et al. shows just that -...about a month and a half, without the need for complicated cloning of homology arms. MMEJ: An Introduction...required by MMEJ can easily be added via PCR amplification. Given the inefficiency of HDR for knock-ins... and 3’ junctions, respectively. These results indicate that PITCh is a robust method for genomic insertion...lies in the diversity of nucleases and their applications. It will be interesting to see what new editing...PMID: 26678082. Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015...633. PMID: 26456817. Find plasmids from this publication at Addgene. 3. Zhang, Chi, Xiuhua Meng, Xiaolei... -
Addgene Stands with the Scientific Community
TypeBlog Post...transformative biological discoveries and life-saving medical breakthroughs and undermine critical research ...Addgene as a trusted partner for archiving their biological materials during funding transitions. Enabling...operating model allows researchers to access authenticated materials at a fraction of the cost of creating...be listed as direct costs from a CRO in grant applications, addressing the new funding structures and indirect...private partners to create additional pathways for critical scientific work to continue. Moving Forward ... -
Make a Splash: Notions of Scientific Impact Are Evolving
TypeBlog Post... citations for recent articles. Of course, a publication in Science or Nature is always nice, but in the... lesser-known journal as it is in a top-tier publication. Accessibility is surely also behind the increase...increase in citations for older publications. (As an interesting aside, Nature recently reported that the...in a solution.) But are citations and journal publications really the best way to measure impact? As that...papers detailing Nobel Prize-winning research typically weren’t among the most cited. And, as scientific...and what it’s made of. Increasingly, journal publications alone aren’t enough. Scientists are sharing ...consider and quantify the impact of scientific publications. Ventures like Impact Story seek to find ways... -
Plasmid Preps: Different Purity, Different Quantities, Different Uses
TypeBlog Post... and chemical contaminants that have been used during the purification procedure. Good indicators of DNA... is necessary for bacterial transformation of chemically and electrocompetent cells is minimal (picogram...nm should fall ~1.8 - 2 and give scientists an indication of purity versus protein contaminants. The 260...nm ratios should fall ~2 - 2.2, and provide an indication of purity versus chaotropic agents as guanidine...addition to being free of any nucleic acid and chemical contaminants, these DNA samples must also be entirely... -
Inntags: Innovative Protein Epitope Tagging
TypeBlog Post...immunoprecipitation applications, but none of the traditional tags were suitable for both applications. To prove...from non-vertebrate systems. Preliminary testing indicated that inntags IT5, IT6, and IT10 expressed well...and the chaperone Ydj1, both of which display sophisticated interactions with multiple other proteins. Again...detrimental effects on cell size and growth rate, indicating that they compromised protein-protein interactions...These tags are easy to use in multiple common applications. Due to their limited effects on native protein...particularly in single-molecule or high-throughput applications. Perhaps IT5 and IT6 will even replace our older... -
Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2: Reprogramming and Transdifferentiation
TypeBlog Post...several ways. Below is a table listing some of critical differences (adapted from Zhou and Melton, 2008...this process. Moreover, it is much easier to genetically modify cells during the reprogramming process...This opens up a wide range of possibilities in clinical situations. In cases where the objective is to...Kushi) Mukherjee is the Editor of Trends in Pharmacological Sciences, a Cell Press reviews journal. She...joined Cell Press to pursue a career in science communication and publishing after completing her postdoctoral...from Massachusetts General Hospital and Harvard Medical School. Connect with her on LinkedIn @ https://...PMCID: PMC2854424. 23. Burridge, P.W., et al., Chemically defined generation of human cardiomyocytes. Nat... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog Post... the bacteria were injected, limiting their applicability. They also did not scatter ultrasound nonlinearly...contrast in E. coli. We deleted an unannotated hypothetical protein, improving expression, and named this...promoter pBAD and a low-copy number origin of replication (p15A) worked best under a wide range of inducer...signal inside the tumors was overlaid with the anatomical B-mode images in greyscale. ARG expression...of the mARG plamsids into HEK293T cells using a chemical transfection reagent (e.g. PEI-MAX). Simply cotransfect...-specific ultrasound signal is in hot and the anatomical B-mode images are in greyscale. (scalebar = 2mm...authors: Marjorie Buss is a PhD student studying chemical engineering. Mengtong "Tom" Duan is a PhD student... -
Hot Plasmids - August 2020
TypeBlog Post...Fluorescent tools for mammalian cells Nanobody purification CRISPRi and CRISPRa system for hPSCs The CRISPR...activity with a fluorescent protein--no affinity purification required! The team used other molecular techniques...compartments in mammalian cells By Angela Abitua Genetically encoded fluorescent proteins are often used to...collaborators have an ongoing effort to improve genetically encoded fluorescent markers for labeling specific...authors generated and described several improved genetically encoded fluorescent markers tagged with mTurquoise2...beads. Workflow of nanobody expression and purification. Iwaki et al., Protein Expr Purif, 2020. ...dCas9 and the gRNA. This expression can vary significantly depending on transgene design and delivery. ...