Skip to main content

We narrowed to 517 results for: Trac

Showing: 161 - 200 of 517 results
  1. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ..., the DNA of interest, and cell extract.    Graphical Abstract: Schematic representation of cell-free...the expression of fluorescent proteins and oxytetracycline enzymes. Next, this system was used to recreate...
  2. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    ...same frequency, we reasoned that, following DNA extraction, the [+] and [-] strands might naturally associate...restriction enzyme based methods, we showed that the DNA extract is a heterogeneous mix of single-stranded and double-stranded...species. While not all of the molecules in the DNA extract associate with complementary strands, enough double-stranded...
  3. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

    Type
    Blog Post
    ...Latin for 'it'), as “a generic term for any extrachromsomal hereditary determinant”. His proposal, however...1960s when scientists began to study other extrachromosomal particles, particularly Resistance or R-factors...experiment. Dr. Cohen and colleagues treated a tetracycline resistant plasmid, pSC101, and a newly developed...
  4. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ...cell!)—ATP. There are biosensors for intracellular, extracellular, and even the ADP:ATP ratio. Most of...sensors developed for it. Calcium triggers muscle contraction, and calcium channels are activated for neuronal...
  5. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...instead of by another method? Viral delivery is attractive due to the ease of transduction in difficult-...human transcription factors, barcoded libraries to trace cell lineage, and so much more! Libraries allow ...as delivery mechanisms for gene therapy. The tractability of AAV, low immunogenicity, and cell-specific...
  6. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...RNA cargo into their targets makes them attractive and tractable tools for genome engineering. Scientists... into an organism. This feature makes them an attractive tool for human subject delivery, as immunogenicity...
  7. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

    Type
    Blog Post
    ... to perform in vivo optogenetic experiments. Intracranial injections of AAV are commonly used to deliver...neurons and mouse brain slices collected following intracranial injection of an AAV expressing a ChRger. In ...nervous system which eliminates the need for intracranial injections. When packaged in PHP.eB, ChRgers...
  8. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    ...the lab observed abnormal cell migration and contraction. The lab hypothesized that this is abnormally...identify whether any rescue cell migration and contraction defects. The lab also plans to use CRISPR plasmids...intensity-based signaling biosensors to monitor intracellular signaling,” says Hébert. Previous research has...
  9. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    ...of using social networking to start, build, and track professional relationships. Here are my seven best... for example). This is also a great way to keep track of how you met the connection, as these notes are...my paper resume). It’s also a great way to keep track of past accomplishments. Follow all the rules of...
  10. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...localization in cells, or even single-particle tracking of the bound complex. There are so many FPs available...TagBiFC technique allows long-term single-molecule tracking of protein-protein interactions in living cells... can even be bright enough for single-molecule tracking in live cells.  Controls  When designing experiments...
  11. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...Epigenetic engineering with single-pot assembly Tracking cell-cell interactions New auxin inducible degron...protocols.io.brgcm3sw     New system developed to track cell-cell interactions By Jennifer Tsang GFP-based...human body and G-baToN provides versibility in tracking and modulating the interactions between cells....
  12. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...BioID or Split-BioID, cells are lysed, protein extracted, and labeled protein interactors purified via ...speed of labeling. BioID requires 6-24 hours for tetracycline induced expression of the BirA* fusion protein...Split-BioID, Schopp et al allowed 24 hours for tetracycline induced expression and biotinylation of interacting...
  13. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...Cellular activities like action potentials, muscle contractions, and the control of voltage gated ion channels...insufficient level. The heartbeat – a muscle contraction – is directly related to the electrical impulse...and emit signal from. This specific example is attractive due to the large number of sheer molecules and...
  14. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...ultrafugation Overview: Iodixanol gradient ultracentrifugation uses a gradient of different concentrations...during AAV production is overlaid. Following ultracentrifugation, the AAV-containing 40% fraction is collected...day. Refer to the AAV Iodixanol Gradient Ultracentrifugation Protocol for more details. Pro tips Watch...
  15. Plasmids 101: Protein Expression

    Type
    Blog Post
    ...protein of interest. Modified proteins can then be extracted from lysed cells through the use of protein tags...forming inclusion bodies that are difficult to extract without harsh reagents and patience. Plant expression... of complex post-translational modifications. Extraction and purification of recombinant proteins from...
  16. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...particles (Figure 1). Briefly, packaged DNA is extracted from purified, DNase-treated AAV and submitted...with AAV-68544 to validate our method. We then extracted the DNA and processed the mixed sample in our ...Adeno-Associated Viral Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678 . https://doi.org...
  17. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ... method (Tanenbaum et al., 2014). The sgRNA or tracrRNA sequences can also be modified to include RNA ...transcription factors. The Root and Doench labs utilized tracrRNAs with two PP7 stem loops to recruit additional ...the targeted region (Sanson et al., 2018) . This tracrRNA was combined with the sgRNA expression vector ...
  18. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...kb), and they remain popular for targeting and tracing cells in the brain (Figure 1). 2nd generation lentiviral...been used for complex applications like lineage tracing in heterogeneous tumors. Lentiviral vectors are...In-vivo RGB marking and multicolour single-cell tracking in the adult brain. Gomez-Nicola D, Riecken K,...
  19. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...biosensors has been designed to detect a variety of intracellular conditions, including ion (such as Ca2+) concentrations...Because of its stability, GFP can be used in lineage tracking capacities in cell fate studies. It can also be...strand break repair, and to label pathogenic intracellular microbes to visualize host/pathogen interactions...
  20. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ...a multigene construct. Figure exemplifies the extraction and use of a coding sequence of interestCDS Level...high-throughput methods to streamline your plasmid extractions. Use this protocol shared by Dr. Rob Hurt for...can check colonies either by PCR, by directly extracting and sequencing the plasmid, or both! We strongly...
  21. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ...certain number of days and tracking average transit time for each courier we contracted with so we can always...Culture Collections Lentiviral Plasmids Tetracyline Inducible Plasmids Empty Backbones Bacterial ...
  22. Hot Plasmids - November 2023

    Type
    Blog Post
    ...cargo Optogenetic proximity labeling and TransitID Tracking cellular history with protein ticker tape Teaching... bioRxiv – doi: 10.1101/2023.02.07.527548.    Tracking cellular history with protein ticker tape By Brian...et al. (2023). Time-tagged ticker tapes for intracellular recordings. Nature Biotechnology, 41(5), 631...
  23. Hot Plasmids - August 2020

    Type
    Blog Post
    ...cytosol (instead of on the yeast extracellular surface) which enables tracking of protease activity with a ...reporter. With this system, they could quantify and track cells expressing both dCas9 and gRNAs. The...
  24. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...scientists are paid above this threshold or for tracking the hours that these scientists work and paying...across academia). Increased postdoc wages, overtime tracking? What are institutions doing? Institutions are...their salary raised 101% if hours are not to be tracked. University of Buffalo and Ohio State both have...
  25. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...General expression mRNA Tetracycline response element promoter Inducible with Tetracyline or its derivatives...promoter with low basal activity and several tetracycline operators. Transcription can be turned on or...
  26. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...like lamellipodia or filopodia formation, cell contraction, or other morphological changes, depending on...tools to control formation of lamellipodia and contractile stress fibers, and recently developed a full ...selection in mammalian cells used here could be an attractive strategy for engineering other enzymes. Conclusion...
  27. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ... signal is boosted and enables single molecule tracking in living cells. The HA frankenbody is an HA tag...expressed on T cells to make CAR T cells. The extracellular scFv portion of the receptors recognizes an ...an antigen of interest while the intracellular T cell receptor portion of the CAR facilitates signal transduction...
  28. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...thrive in laboratory settings. One of its most attractive features is its stability and easily obtainable...ability to regenerate, the axolotl has several attractive characteristics that make them an ideal model...for human disease research. One of their most attractive features are their eggs. Frog eggs are large,...
  29. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ...useful to avoid such procrastination: Remove distractions: Probably the easiest way to do this is to leave...internet and all it’s enticements are far too distracting. (As a side note, I’d recommend the book, “Bored...more fantastic reasons to remove yourself from distractions every once in awhile.) Reframe the task in a...
  30. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ...therapeutics, which is also gaining a lot of new traction lately (1, 2), depends on knowledge of the structure...storage. After overnight growth, scrape up, pool and extract plasmid DNA directly from the colonies. Transform...solution containing your favourite bacterial protein extraction reagent (BPER from Thermo works well) with some...
  31. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...the sensitive FLEx-split TVA receptor used for tracing neural pathways was 8.5 x 10e10 GC/mL. The lot ...Adeno-Associated Viral Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678. https://doi.org...Jin L, Lea NE, Wickersham IR (2020) Monosynaptic Tracing Success Depends Critically on Helper Virus Concentrations...
  32. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ...culture through various sources that are difficult to trace. These include the laboratory personnel, the serum...mycoplasmas, contamination by other bacteria or extracellular fluorescent signals produced by these bacteria...fluoroquinolone ciprofloxacin, ciprobay, zagam, baytril, tetracycline, etc. are available for mycoplasma removal from...
  33. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...microglia. This property makes the lentivirus an attractive choice for a gene transfer vector. Lentiviral...used inducible lentiviral vector system is the tetracycline (Tet)-regulated system. You can choose either...promoter in your transfer vector. In the absence of tetracycline or a derivative such as Doxycycline (Dox), the...
  34. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    .... Other dyes may prove challenging to stain intracellular compartments in in vivo model systems, so extra...green, red, and far-red dyes is important for intracellular fluorescence live imaging ​​(Hayashi-Takanaka...fission, fusion, biogenesis, and mitophagy, can be tracked using dyes like Rhodamine 123 (R123) (Liu et al...
Showing: 161 - 200 of 517 results