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Showing: 121 - 140 of 502 results
  1. The Challenges of Cell Culture

    Type
    Blog Post
    ...records allows you to track a cell line’s age and monitor any changes. Your tracking system could be as ...just a few publications which have since been retracted due to cell line contamination: Epithelial Mesenchymal...colleague. Time and money are wasted, papers are retracted and there is a loss of reputation that can impede... good recordkeeping, you need to have a way of tracking how many passages the cells have undergone and...
  2. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ... so, the WHO recommends one strategy: testing, tracking, and social distancing. With countries like South...block and 62°C heat block Approach: Viral RNA is extracted and subject to isothermal amplification by primers...directly using saliva without having to perform RNA extraction but that may work only if there is a high enough...water bath and 42°C water bath Approach: RNA is extracted from a sample and subject to isothermal recombinase...detect SARS-CoV-2 in the samples. First, RNA is extracted and subject to reverse transcription. This is ...
  3. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...has become very useful in lineage tracing; “Zebrabow” was used to trace the development of the corneal epithelium... also been applied to cell marking and lineage tracing in other systems, such as zebrafish and Drosophila...Zebrabow: multispectral cell labeling for cell tracing and lineage analysis in zebrafish. Pan YA, Freundlich...dev.094631. PubMed. Multiplex cell and lineage tracking with combinatorial labels. Loulier K, Barry R,...
  4. Finding Your Perfect Job After University

    Type
    Blog Post
    ...companies which are reliant on contracts with clients is that, when those contracts fall through or are cancelled...(SOP’s) and every item I tested was constantly tracked with barcodes and spreadsheets. It was great to...primary purpose of the company was to fulfill contracts, my work sometimes felt rushed and the focus always...company; new staff hired in anticipation of the contract suddenly have less work to do than was anticipated...
  5. Important Considerations When Using AAVs

    Type
    Blog Post
    ...feature of AAVs as a new tool for neuroanatomical tracing (11). With the ability to produce neuron specific...showing a very specific phenotype. Previous neural tracing methods used viruses that were potentially toxic...available, AAVs could be much better tools for neural tracing in very specific cell populations. Find Ready-to-Use...finger nucleases, and many others (12). The most attractive potential of AAVs is their ability to be used...et al. "A fourth generation of neuroanatomical tracing techniques: exploiting the offspring of genetic...
  6. Plasmid Preps: Different Purity, Different Quantities, Different Uses

    Type
    Blog Post
    ... cells is minimal (picogram range). It can be extracted from small bacterial cultures (i.e. 2-3 ml) with...cultures and commercial kits you will adopt for the extraction. Restriction enzyme based approaches can require...and guanidine hydrochloride used during plasmid extraction. Figure 2: Use cloning grade DNA for direct... technique must be larger scale, to allow the extraction of enough plasmid to last for multiple experiments...
  7. Addgene's Top Blog Posts from 2020

    Type
    Blog Post
    ...complement our RNA Extraction Without A Kit blog post, we've compiled five methods for extracting DNA without...that cause plasmids to be incompatible. 4. RNA Extraction Without A Kit by Leah Schiesow We added to our...it, wrote these tips for working with RNA and extracting RNA without a kit. 3. SARS-CoV-2/COVID-19 Detection...
  8. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...cells. They bind to proteins on the surface of extracellular pathogens such as parasites or microbes, or ...from the animal is drawn and the antibodies are extracted. These antibodies recognize a variety of epitopes...Hybridomas are formed by fusing B cells that have been extracted from animals injected with protein antigen to ...presence of protein within a sample, you’ll want to extract your protein of interest from the mixture. Antibodies...permeabilized to allow antibodies to bind to intracellular proteins. The fluorescence signal is detected...
  9. Three Tips to Organize Your Lab Notebooks in the New Year

    Type
    Blog Post
    ... deadline again. Organize you experiments, keep track of upcoming grant deadlines, and sync everything...Though electronic lab notebooks (ELNs) have gained traction with pharmaceutical companies, academic labs have...allow you to control who can edit the documents and track the revision history. Now you’ll be able to make... knowing the information is backed up and edit tracking will help you identify when others have added ...
  10. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...the intensity of fluorescence signal and enables tracking of single molecules within living cells without...thereby creating a single-molecule reporter of intracellular processes. Multimerization via the SunTag system....3147. Bioluminescent pH sensor Understanding intracellular pH regulation is important as pH regulation ...to the cell cycle. Good tools for measuring intracellular pH are required in order to determine the relationships.../Cas9 in genome-engineering. One particularly attractive application involves using dCas9-activator fusion... compartment (e.g. the plasma membrane, the extracellular space, or the lysosomes) can take many paths...synchronous release of the reporters which can then be tracked along the secretory pathway (Figure B). The RUSH...
  11. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    ...bound to the intracellular surface of the GPCR. Binding of a ligand to the extracellular surface of the...environmental conditions, cells must be able to detect extracellular stimuli. One way they do so is through G protein-coupled...Slocum ST, Che T, Gavin AC, McCorvy JD, Roth BL, Strachan RT (2020) TRUPATH, an open-source biosensor platform...
  12. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    ... calculate the amount of bound ligand (after subtracting out non-specific binding). Historically, glutathione-S-transferase...subsequently purify the fusion protein from a bacterial extract. The MBP tag can be removed using Xa protease. ...successfully used this plasmid to biotinylate the extracellular domain of the erythropoietin receptor and subsequently...biotin (50X) because 5 mM is barely soluble at RT. Track expression and purification of biotinylated protein...
  13. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    ...barcodes within the gRNA sequences. This allowed tracing of the performance of each gRNA multiple times ...protein phosphatase that responds to rising intracellular calcium levels within the cell. When levels ...required for virulence in pathogenic fungi, it is an attractive drug target. Calcineurin reporters allow scientists... wild type (top panels) and cells with high intracellular Ca2+ (bottom panels) equipped with destabilized...
  14. The MTA at Addgene

    Type
    Blog Post
    ...centralized tracking when biological reagents are shared — both at what time and with whom. Tracking these ...read on!  What is the MTA?  An MTA is simply a contract or agreement that states the terms of use for ...meaningful progress depends on things being organized, tracked, and clearly communicated. And that’s where MTAs...
  15. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    ...autophagosomal membranes, so tagging it with GFP allows for tracking the localization or accumulation of pre-autophagosomal...an autophagy-independent manner. Additionally, tracking Atg8p or LC3 doesn’t provide information about...easier to study the mechanisms behind autophagy by tracking what’s being transported to the yeast vacuole ...vacuole (i.e. cytosol, mitochondria), rather than tracking generic autophagy markers. It’s particularly useful...
  16. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ... the crRNA or the tracrRNA. Of course, in a single guide system (crRNA and tracrRNA as a single molecule...region associates with the trans-activating crRNA (tracrRNA) which interacts with the Cas9 protein. These ...you to visualize transfection efficiency and even track localization in cellular experiments. There are ...
  17. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ...concentrated by a variety of methods such as ultracentrifugation, centrifugation on a sucrose gradient, spin...number of 10 cm2 dishes. While this option is attractive because the same procedure for a single 10 cm2...relatively inexpensive and does not require an ultracentrifuge, PEG will precipitate all proteins in a solution...high-titer lentivirus preparations without ultracentrifugation. Sci Rep. 2015; 5:13875. PubMed PMID: 26348152...
  18. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...behind from a preceding animal (ex: urine) can be distracting. If it is not cleaned, animals tested towards...experiment series would behave very differently (distracted) from the ones tested at the beginning. Variability...Both AAV and lentiviral vectors are used for intracerebral injections. AAV particles are smaller and may...liquor of the cerebral ventricles (ICV, intracerebroventricular) to minimize tissue damage; however, the...
  19. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...better access to intracellular targets. If your target is already on the extracellular side of the membrane...is suitable for antibodies that bind to the extracellular portion of a protein. B) Permeabilizing with...antibodies into the cell where they could bind to intracellular epitopes. C) Permeabilizing with a non-ionic...
Showing: 121 - 140 of 502 results