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We narrowed to 921 results for: Tro;

Showing: 451 - 480 of 921 results

  1. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    ...antibody, you’ll want to use the same positive control across experiments to be aware of signal strength...you’ll want to be sure to choose the appropriate controls to be sure the staining you are seeing is real...real. It’s important to include a negative control using serum from the same species your polyclonal antibody...antibody was generated in or a polyclonal isotype control, to account for nonspecific binding. Now that you...

  2. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ...localization. Surprisingly our observations at the electron microscope showed us that, in monkey neurons, ...of the haemagglutinin of the influenza virus, introduced to the field of fusion proteins more than thirty... of interest the membrane.   Figure 1: Electron microscope images showing ultrastructural localization...protein of interest, an alternative could be bicistronic expression of the fluorescent protein with the...

  3. Plasmids 101: Cre-lox

    Type
    Blog Post
    ...AAV (AAV-pgk-Cre) has been used to successfully introduce Cre into cells of interest. What if I need two...of using loxP sites is the inability to tightly control which loxP sites recombine if more than two are...allowing for two separate recombination events to be controlled in parallel. Have you used the Cre-lox system...PubMed Central PMCID: PMC49604. 4. Independent control of immunoglobulin switch recombination at individual...

  4. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ...chemogenetics? Chemogenetics relies on genetically introducing engineered receptors that are only activated ...Expression Using chemogenetics requires successfully introducing receptors to target cells. Targeted and specific...mitigated by using low doses and running appropriate controls (Jendryka et al., 2019), they have resulted in...channel, the PSAM acts as a gating mechanism to control ion flux. The system allows you to directly manipulate...

  5. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...live cells.  Controls  When designing experiments, it’s important to include proper controls to assess the...quantitative readout of the interaction in vivo or in vitro, and can be used for measuring protein expression... p53 tetramerization in response to DNA damage controls the p53 response. Gaglia G, Lahav G. Mol Syst ...

  6. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...can be exploited to either inactivate a gene or introduce a desired genetic alteration. Listen to Our Podcast...necessary gRNAs, but it would not be possible to control which gRNA becomes complexed with which variant...represents a straightforward means of exerting tight control over Cas9 nuclease activity while eliminating the...guides targeting one set of genetic loci can be introduced together with truncated gRNAs targeting a different...

  7. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ... proteins. The Seeger lab developed a robust in-vitro protocol that enables rapid selection of synthetic...sybodies that hypothetically contain ones that bind strongly to your target protein. The Seeger lab designed...selection step involves using a commercial kit for in-vitro translation of the mRNA libraries. Each mRNA molecule...enrichment step for identifying sybodies that bind strongly and specifically to the protein of interest. The...

  8. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...advantages degron tags can offer over other protein control methods and compare the commonly available types...etc. Firstly, they are reversible mechanisms of control. Protein degradation can be induced by the addition... Tsien RY, Lin MZ. Tunable and reversible drug control of protein production via a self-excising degron...  Susanna Stroik...

  9. Of Myc and Men

    Type
    Blog Post
    ...of interest to better study them in vivo and in vitro, especially in the mid-late 20th century with the...reticulum localization, Munro and Pelham casually introduced the Myc-tag (Munro, et al. 1986; Munro and Pelham... and the Myc-tag today Since Munro and Pelham introduced the Myc tag to the world, it has become one of...this antibody works and allow users to better troubleshoot and design experiments. It turns out that even...

  10. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...experiments attempting to build gene drives in tropical mosquito vectors of diseases such as malaria and...then, using multiple confinement strategies is strongly advised. The effects are multiplicative, and there.... Site-specific selfish genes as tools for the control and genetic engineering of natural populations....Fitz-Earle, M., Holm, D. G. & Suzuki, D. T. Genetic Control of Insect Populations: I. Cage Studies of Chromosome...

  11. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...and would avoid genomic off-targets or indels introduced through non-homologous end joining (NHEJ). Designing...recapitulate or rescue known pathogenic alleles, or introduce a premature stop codon to render an RNA nonfunctional...also adjusted the structure of the guide RNA by introducing a C opposite the target A to be edited. This ...off-target edits around the targeted nucleotide. Introduction of guanine-mismatches in the guide across from...

  12. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ...for Laboratory Automation and Screening (SLAS) Electronic Laboratory Neighborhood.   Work/life balance....possible to bring all aspects of our lives under control? Balance is a myth and should not even be the real... don’t have children, so my life will be under control.” Think again. Maybe you’ll need to care for an...to do most tasks – try to resist the impulse to control by insisting things always be done your way. Be...

  13. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ...founder of Research Space. There are many types of electronic lab notebooks (ELNs), each with its own pros ... virtue of liberating data from paper into an electronic environment and hence making it searchable and... of Research Space, which provides the RSpace electronic lab notebook. Rory is based in Cambridge, Massachusetts...on the Addgene blog Find tips on choosing an electronic lab notebook Watch a livestream of the Minisymposium...

  14. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...and selection for targets can be done largely in vitro; Easy and reproducible commercial synthesis (those..., Akhtar and Marty, Jean L. “Aptamer based electrochemical sensors for emerging environmental pollutants...systems using immobilized aptamers.” Biosens Bioelectron. 2011 May 15;26(9):3725-36. PubMed PMID: 21419619...FRET in Your Experiments Learn more about GFP Controlling Protein Activity with GFP Resources at Addgene.org...

  15. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...social sciences go uncited. Creative titles and controversial topics can garner some attention, though in ...Journal of Number Theory (1,519 views), and Gastroenterology (1,046 views) rounded out the top four average...infections. The video abstracts provide a brief introduction to the research and, after watching the video...links to the videos, keeping the copyright and control over the videos with the researcher and the host...

  16. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ... Martin et al., 2018). The ACE reporter is a bicistronic construct that consists of a mutated mCherry ...eGFP. To create the inactive mCherry, the lab introduced a 43 base-pair insertion that disrupts fluorescence...use across different cell types and systems. To introduce flexibility, the Dow lab created a base editing... optimized cytidine base editor. The lab then introduced an sgRNA targeting the GFPACG to the cells. As...

  17. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    ...reproducible assay. That information can make you a much stronger candidate!  Additional uses The cover letter is...job with a lot of file management that asks for strong organizational skills. You might think, “I’d be...letter The opening paragraph is the place to introduce yourself and your background. It’s a great place...and is important for the job]. Notice that the intros above allow you to highlight what specific skills...

  18. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...protocols therefore differ from library to library. We strongly recommend following the protocols provided by ...a low multiplicity of infection (MOI). MOI is controlled by viral titer, so you’ll need to measure your...Addgene uses a physical titer (via ddPCR) to quality control our viral preps, we recommend determining the functional...transduction protocol you can follow and a troubleshooting blog should you run into any issues.  Keep ...

  19. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...overexpression vector rather than going through all the troubleshooting required for Cas9-based activation. CRISPRa...Dow D, Buser-Doepner C, Bertotti A, Trusolino L, Stronach EA, Saez-Rodriguez J, Yusa K, Garnett MJ (2019...Weissman JS (2014) Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Cell 159:647...2015) A light-inducible CRISPR-Cas9 system for control of endogenous gene activation. Nat Chem Biol 11...

  20. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...neither the aptamer nor the small molecule are strongly fluorescent but the binding of an aptamer to its...plasmids encoding aptamer-tagged RNA and later introducing the small molecule into the cell. The fluorescence...metabolites, leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches...then identified through microfluidic-assisted in vitro compartmentalisation for their high binding affinity...

  21. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...Level 1 microbe makes it a perfect candidate for introductory teaching and research environments (Idalia &...study human cardiovascular disease, specifically stroke and hypertension (Iannaccone et al., 2009). Rodents...Resource, a breeding facility for X. laevis and X. tropicalis, the two most commonly used species of frog. ... study human disease-relevant genes in such a controlled environment displays how essential the frog is...

  22. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    ...reviewer’s concerns, you are making your hypothesis stronger. When you share data at a conference poster session... can frequently be traced back to one of a few trouble spots:  Communication (or lack thereof): Information...Empathizing with people, especially people with whom you strongly disagree, is a critical component of conflict ...tools I’m mentioning here, this may be the most controversial, so I would like to qualify it. If you are attempting...

  23. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...while shRNA and siRNA are synthetic constructs introduced through external methods. shRNA processing Following...expressed constitutively, either in lentiviral or retroviral backbones. On some occasions, shRNAs are transiently...transient transfections and are handy for quick in vitro experiments. shRNA plasmids typically use RNA polymerase...common promoters are U6, H1, and 7SK. These are strong promoters with no tissue specificity, so in some...

  24. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...most important ways to spatially and temporally control gene expression, especially in in vivo models, ...Golgi staining, injection of diffusible labels, or electron microscopy of stained sections, were limited due...functionally similar to Cre-based Brainbow, but is controlled by FLP/FRT recombinase. When placed under different.... The use of AAV provides spatial and temporal control without the need for germline modification, and...

  25. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...as the insertion of a fluorescent tag or the introduction of a specific mutation, one generally relies...one but two rounds of single cell cloning – as a control, one should revert the edit back to the original...technologies offer an alternative to HDR for introducing edits. The same locational constraints are even...the method of producing the guide (synthetic, in vitro transcription, or lentiviral delivery) can affect...

  26. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    ...priorities and needs, but sometimes it just takes introspective thought on what’s worked for you and not worked...reputation? If you are choosing a training lab, I strongly suggest you complete a work rotation in the lab... it could also be a sign that the lab head has trouble attracting scientists. To get a better idea of ...and non-academic career paths. I can’t say this strongly enough - don’t choose a mentor that sees you only...

  27. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ...formation in the genome and are inefficient at introducing point mutations. In 2016, the researchers at ... Broad Institute envisioned a new strategy to introduce point mutations in the genome. They fused a catalytically...base editors (CBE) use a cytidine deaminase to introduce a C→T change at precise locations in the genome...his super giant albino gorilla fighting off a monstrous wolf and crocodile, that were “affected by CRISPR...

  28. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...researcher in the University of Michigan Department of Astronomy. She studies how planets form around young stars...community of science supporters is much larger and stronger that I expected. Jennifer Stancill, Ph.D., is ... an agenda that halts current data collection, destroys existing data, and guts future funding.I marched...march read "You know things are serious when the introverts arrive." I couldn’t watch the denial of facts...

  30. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...initiate our MTA process and perform standard quality control but will not make the plasmids available until:...Continue to Next Step". As part of our quality control process, we will sequence the full plasmid and ...recombination in bacteria) such as lentiviral, retroviral, and AAV plasmids, we recommend the NEB Stable...received them and they will undergo our quality control process to verify your plasmids. Our friendly Addgene...
Showing: 451 - 480 of 921 results