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We narrowed to 1,010 results for: CAL;

Showing: 201 - 220 of 1010 results

  1. Five Popular Model Organisms

    Type
    Blog Post
    ...mice at a time. Because mice are more genetically and physiologically similar to humans than many other model...study calcium signaling in the entire adult brain, using a plasmid that expresses a genetically encoded... of mice as model organisms is that they are genetically tractable. Mice can be easily manipulated with...also relatively straightforward to manipulate genetically. Scientists can make a new line of flies in only...reasons above but because they actually share many biological properties and processes with our own cells. ...important experimental model in the study of neurological diseases, such as Parkinson’s disease (PD). ...embryonic development to adulthood. This ability to genetically manipulate and visualize cells and tissues allows...

  2. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    ...have been typically optimised for immortal/genetically-corrupt cell lines, lack scalability, and have ...alone in the cellular milieu. It is, therefore, critical to understand how proteins interact with one another... another (or with DNA) to perform diverse biochemical tasks in the cell. One of the most popular approaches... to tag the protein of interest with a small biochemical “epitope tag” such that these tagged proteins...epitope tagging in mammalian stem cells. For easy scalability to the 96-well format, we developed a “TAG-IN... purchased from various other suppliers with a typical turnaround time of 5-7 days. The crRNA and tracrRNA...optimized to achieve high knock-in efficiency. To calculate the tagging efficiency in the transfected cells...

  3. Deep Dive: qPCR

    Type
    Blog Post
    ...which aims to increase your understanding and technical proficiency with common applications - now let... differences of twofold or greater with good statistical significance (Taylor, 2017). With the right conditions...appropriate choice (Panina, 2018). Look for studies specifically aimed at determining the best options for a ...of triplicates to understand variation due to technical error.  Plate readouts can be affected by plate...sets and most machines have a protocol that automatically performs the normalization step.  Machines Most...when using fluorescent probe-based chemistry - theoretically, up to five or six reactions in a single well...and sets.   🔥Hot Tip🔥 The ABI machines automatically record fluorescent data on all channels - helpful...

  4. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...editors (CBEs) and adenine base editors (ABEs) chemically modify target bases without breaking the DNA ...one-ring pyrimidine base. This is a far more complex chemical task, not a job for a single enzyme. Base ...editing works in general. Base editors use either catalytically dead Cas9 (dCas9) or a Cas9 nickase (nCas9) ...adjacent DNA to create a region of separate strands called an R-loop (Figure 2). This allows the guide RNA...terminology when discussing base editors, like calling it the “edited strand.” Uracil excision repair... an engineered adenosine deaminase capable of chemically converting adenosine (A) to inosine (I, treated...converting the C base to uracil (U). This is the same chemical reaction performed by CBEs. However, most CBEs...

  5. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...the sandwich and typically takes less than an hour. However large proteins typically do not transfer as... boiling in the presence of a reducing agent, typically containing thiols, to cleave disulfide bonds. ...gel on top of the resolving gel. A stacking gel typically has a different ionic strength and lower pH and...nitrocellulose or polyvinylidene difluoride (PVDF) in a step called protein transfer. While both nitrocellulose and...are “sticky” and have the potential to bind nonspecifically to the antibodies used for staining. To address... the controls listed on the vendor’s website. Typically, the vendor will demonstrate that their antibody...binds to endogenously expressed protein at physiologically relevant levels. Some vendors will also test...

  6. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...ways to monitor cell, organelle, and protein localization. For instance, you might dye your cells and ...timers, add an entire new dimension to monitoring localization; using FP timers, researchers can look at a ...single image of a cell and understand how protein localization changes over time.Hallmarks of fluorescent protein...time but independent of protein concentration. Calibration curves correlating this ratio to the total expression...in whole cells or as a pool of fusion proteins localized to a specific region of the cell. For example,...fluorescence falls within the linear window for the theoretical timer shown in Figure 1B, you can use the equation...temperature, and pH, but timer activity can be calibrated to these different conditions, enabling the use...

  7. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ...fluorescence microscopy vs. confocal microscopy In confocal microscopes, optical sectioning of a specimen...sheet, typically 2-6  microns, illuminates only one thin plane of the sample surrounding the focal plane...Lee and Pantelis Tsoulfas of the Department of Neurological Surgery at the University of Miami. The beginning...expressing eGFP under the human ubiquitin C promoter. Anatomical abbreviations: Gr= Gracile tract, Cu= Cuneate...originally named orthogonal-plane fluorescence optical sectioning (OPFOS) (2). Arne Voie, David Burns ...illumination (i.e. light sheet).  This is in contrast to confocal laser microscopy where the laser and the reflected..., the photomultipliers used to detect light in confocal microscopes are less efficient than the modern...

  8. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...visualize the localization of the FP (fused to the protein of interest), thus tracking the localization of that.... The most commonly used are GECIs - genetically encoded calcium indicators (e.g. GCaMP). Others include...capacity. Many of these FPs are fluorescent when ectopically expressed in most organisms. Furthermore, fusing...fluorescence. Therefore, FPs are used to study many biological questions. The two most common uses are: 1) to...following their activation. The four main FTs are called Slow-FT, Medium-FT, Fast-FT, and mK-GO. Large Stokes...For LSS proteins, the Stokes shift is ≥ 100nm. Specifically, these proteins are excited by UV light or blue...of the split FP. This technique is also is also called Bimolecular Fluorescence Complementation (BiFC)...

  9. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...ratio calculators Lab math calculators These calculators can help you with common calculations performed...lab. Molarity calculator Promega's Biomath Calculators Primer resuspension calculator (sign-in required... Cloning ligation reaction calculators These links can help you calculate how much insert and backbone...required.)  DailyCalcs Science Calculator app for iPhone Reagent and Data Repositories These links can help...assembly, or In-Fusion cloning.  NEB Ligation Calculator: This tool works for any cloning method. You ...

  10. Running for Rare Disease, Running for FOP, Running for AJ

    Type
    Blog Post
    ...Attempts to remove excess bone surgically cause the body to “heal” the surgical site with even more bone that...therapeutic biologic design (ADCs) along with pre-clinical analytical development. His areas of expertise include...protein engineer currently working at Sanofi Pharmaceuticals in Cambridge, MA. When I joined Sanofi/Genzyme...to get in shape. After three kids, I had put on typical middle age weight and “exercise” consisted of taking... learned there was a Genzyme-based running team called Running for Rare Disease that runs on behalf of... But, there is HOPE. FOP is a genetic disease typically caused by a single R206H point mutation on the...growth in mouse models. As this biologic moves into clinical trials, there is new HOPE for AJ. But, as we in...

  11. MXS Chaining

    Type
    Blog Post
    ...BglBricks: A flexible standard for biological part assembly. Journal of biological engineering. 2010;4(1):1. Pubmed... diverse functions, and thus, is used in many biological fields. In synthetic biology for example, high-throughput...elements to generate non-natural tools such as novel biological circuits or sensors. Given the expanding palette... can be used to detect structure and protein localization, as well as cellular processes like gene expression...in-frame to a tethering partner, which directed localization of the fluorophore, and thus enabled labeling...emission maxima Tethering partner Subcellular localization 1 TagBFP 399nm/ 456nm histone 2B (H2B) Chromatin...to generate polycistronic containing multiple identical polyadenylation signals or promoters Gibson et...

  12. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...al.: bowtie2 -f -x HG19_GENOME --local -f -k 10 --very-sensitive-local -L 9 -N 1 -U GRNA_23MERS -S GRNA_HITS.sam...genomic regions that are bound to modified proteins called histones. We can detect modified histones with ... Table 1. Because of DNA’s 3D geometry and the chemical properties of modified histones, a genomic region...maintaining ESCs. So which of these 13,000 regions are critical? One hypothesis: the enhancers! This leads us ...approach for designing an experiment to identify the critical NANOG binding sites: 1. Find Enhancers that have...differentiate Step 3 reveals which NANOG-related genes are critical to stem cell survival. Knowing which genes influence...and therapeutics. We’ll have more to say about clinical applications of CRISPR in the next section. In...

  13. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...fixatives - chemical cross-linkers and organic solvents.  Chemical Cross-Linkers Chemical cross-linkers...artifacts and how to minimize them. FocalPlane. https://focalplane.biologists.com/2020/07/07/fixation-artifacts-and-how-to-minimize-them...-linking cellular proteins. Cross-linking is a chemical reaction that covalently joins two molecules and... confusion. Paraformaldehyde: Many IF protocols call for paraformaldehyde (PFA) as the fixative. PFA ...depolymerizes, producing formaldehyde. Often people will call the resulting solution PFA, but it’s really just...proteins can be lost during fixation with these chemicals, which impacts sample structure and could also... target to be washed away. Furthermore, these chemicals can also alter the structure of proteins, which...

  14. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    ... We’ve also created constructs with containing chemical tags such as the SNAP-tag, Halo-tag, and CLIP-...animals and fix the samples. When you use AAVs, it typically takes a few weeks to achieve optimal expression... Any longer and the cells may start to show morphological abnormalities due to the excessive amount of... levels of XFPs - Reduce the amount of tTA. Paradoxically, too much tTA leads to a reduced expression ...most likely by suppressing transcription. It is critical to express a minimal amount of tTA to achieve ...brain samples, we recommend pretreatment with ScaleCUBIC1 (Susaki et al., 2014), before clearing with SeeDB2...sensory systems focusing on the functional and anatomical maturation of the olfactory system.      Takeshi...

  15. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...get there and more practically, when do you need to be there? A typical biomedical postdoc fellowship ... fail to replicate. Plan experiments logically and strategically to increase your rate of productive results...theJanelia Research Campus of the Howard Hughes Medical Institute. Today's postdoctoral fellows (PFs) face...submitted manuscript in a high-profile journal typically takes 6-12 months depending on the revisions and... to be ready to write by Year 3.5. Thus, it is critical to get an experimental system up and running very...identified a postdoctoral mentor. A good mentor typically writes the grants that fund your research, guides...However, you have finite time and resources. You typically have 4-5 years to make discoveries and publish...

  16. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...circle. Typically, a yellow check mark indicates that a particular parameter was automatically adjusted... The vertical black dotted line represents the cut site. Cutting and error-prone repair typically result... of robust software tools. As a consequence, a critical step in the gene editing workflow - analyzing ...over-looked.  Synthego has developed a new tool called ICE (Inference of CRISPR Edits) to solve this problem...guide RNA sequence you used. The ICE tool will calculate overall editing efficiency and determine the profiles...and interpret data about these types of edits is critical to the CRISPR workflow. For a multiplex sample...steps to learn. For increased flexibility and scalability, the ICE software has two analysis formats: sample...

  17. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    ...dissonance (Kunda, 1990). More specifically, they were engaging in a behaviour called the “biased evaluation ... past my bedtime, poring over my grandparents’ medical textbooks by torchlight under the covers. In high...boasted to me that he never eats “anything with chemicals in it,” leaving me to wonder how he has survived... read a version of the abstract where the real medical conditions had been swapped with made up illnesses...their preconceived ideas about the controversial medical issues. Participants gave significantly more favourable...beliefs even though all three texts were otherwise identical. Likewise, when participants read an abstract ...rate that study as untrustworthy, inaccurate, methodologically flawed, and biased. As a result, the control...

  18. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    ... or poor scalability.  Scientists at Janelia Research Campus and the University of California, Berkeley...across broader physical areas of the brain and enable access to neurons that are physically blocked, such...driven by neurons. By studying how neurons are physically connected, we can begin to understand how the...Therefore, by simply looking at how neurons are physically connected we can begin to understand how the ...region (anterograde tracer, Fig. 1B). Tracers are typically delivered to a particular location and they either...retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses can be injected into the brain and efficiently...neuronal signaling, or manipulating any other genetically controlled function. By using AAVrg in Cre transgenic...

  19. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...protein. The electrical system converts the fluorescence intensity to a voltage pulse, called an event, ...of the population as a whole, scientists gain statistical power in their observations. However, while flow...subcellular analysis such as morphology or subcellular localization studies. For these, alternative methods such...Cells are first resuspended in a pressurized buffer called sheath fluid and transported through tubes or capillaries...in single-file where it is interrogated by the optical system (Figure 1).   Figure 1: Cells expressing...complexity, measured using visible light scatter The optical system has both visible and fluorescent light sources...same direction as it was initially traveling is called forward scatter (FSC) and provides information ...

  20. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...compute using electrical impulses to coordinate emotions, thoughts, and behavior. Historically, scientists...To bring greater control to Acr activity as a biological tool, Niopek and colleagues turned to optogenetics...fusions, and xCas9, and requires no additional chemicals or modifications. This versatility opens up the...scientists study the electrical activity in the brain by inserting electrodes into the brain. But this process...and expensive. Thus scientists have turned to genetically encoded voltage indicators. To expand the voltage...thousands of proteins for one that can report electrical activity via imaging. They took the light-sensitive...light-sensitive protein QuasAr2 and systematically mutated it. Using a robotic picking approach to select cells...
Showing: 201 - 220 of 1010 results