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We narrowed to 797 results for: Tec

Showing: 141 - 160 of 797 results

  1. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    ...CRISPR 101" post. Next-generation sequencing technologies are the gold standard but they remain cost-prohibitive... assays (e.g. Surveyor® or T7E1) for mutation detection. While these methods paved the way for mutation...introduce digital PCR as an emerging validation technology. Digital PCR has several advantages over mismatch... require standard curves, and can theoretically detect a single target molecule in a sample. Digital ... real time PCR. These assays can be designed to detect either a donor sequence integrated through homology-directed... remainder of this post will focus on assays to detect NHEJ. These assays include a forward and reverse...between 200 and 500 ng of purified PCR product. Low detection limits: Digital PCR assays can accurately quantify...

  2. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    ...popular technique in neuroscience and developmental biology. This type of cell labeling technique involves...group developed the original iteration of this technology in 2007. Used to label neurons in the cerebellum...cerebellum of mice, the team appropriately named their technology Brainbow. Brainbow 1.0 used the fluorophores ...construct but do not undergo recombination can be detected by immunostaining of the truncated YFP. In Thy1...CMV-Brainbow construct at the embryonic stage. This technique could be used to target Brainbow to neurons by... promoter. Zebrabow is an extremely versatile technology as recombination can be induced in developing...cells. Tetbow optimizes Brainbow for current technologies Tetbow (Sakaguchi et al., 2018) overcame the...

  3. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...Einstein College of Medicine has recently published techniques to follow translation in living single cells....cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even here, there is the question... symposium seeking inspiration to resolve the technical hurtle of monitoring protein turnover of specific...interesting talk by Prof. Erin M Schuman, a former Caltech researcher, now Director at the Max Planck Institute...Frankfurt am Main, Germany. She presented a novel technology, back then still unpublished, that allowed the...synthesized proteins with FUNCAT Professor Schuman’s technique applies metabolic labeling using a noncanonical...noncanonical amino acid, azidohomoalanine (AHA). In the technique, AHA is incorporated into newly synthesized proteins...

  4. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...Scientist at the Broad Institute, John Doench. CRISPR technology has made it easier than ever both to engineer...is true when designing gRNAs for using CRISPR technology – the “best” gRNA depends an awful lot on what...Gene knockout by NHEJ Gene knockout with CRISPR technology is usually accomplished by Cas9-mediated dsDNA... Created with BioRender.com. Two newer technologies offer an alternative to HDR for introducing ... another target C or A in the window. Another technology, prime editing (reviewed in Anzalone et al., ... although these are still early days for this technology, and the user may need to optimize numerous parameters...CRISPRa (activation) and CRISPRi (inhibition) technologies – a nuclease-dead Cas9 (dCas9) is directed near...

  5. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ... permeabilization techniques — and so it may be necessary to try multiple techniques. For example, we... ability to detect endogenous levels of the target protein. It may only be able to detect the target protein... (or refute) that the antibody is selectively detecting the target-of-interest in your assay and sample-of-interest...must first confirm that a candidate antibody can detect the antigen of interest in the given protocol. ...supportive, the data must show that the antigen being detected in the intended application — here, flow cytometry...conclusively prove the ability of an antibody to detect the antigen of interest when expressed at endogenous... approach, you’re looking for an antibody that detects the target in the parental cell line, but not in...

  6. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    ... and can be used in a multitude of scientific techniques such as molecular cloning or molecular diagnostics...used for an array of different molecular biology techniques and experiments. End point PCR’s goal is to amplify... cloning to create a plasmid of interest, for detecting the presence of an insert or piece of DNA in a...reaction (qPCR) also known as real time PCR is a PCR technique used for measuring a starting DNA concentration...during the PCR reaction cycles is continuously detected via a fluorescent signal. The signal increases...concentration. The cycle in which the unknown DNA is detected compared to the standard curve can be used to ...right. ddPCR uses a water-oil emulsion droplet technology that fractions the PCR reaction sample into approximately...

  7. Common Injection Routes in Mice

    Type
    Blog Post
    ...the mice, these “no needles” techniques may come in handy. These techniques are referred to as “instillation...to conducting mouse experiments. Some of these techniques are what we call “beginner”, or injections that... practice after initial mouse training. Other techniques require more skill and additional training, going...Image created with biorender.com.   Needle techniques Intravenous Intravenous (IV) injections are most...easier of the two, but it actually requires more technical skill to place the needle in the correct vein ...are probably the most advanced needle injection technique. Intrathecal injections are used when the substance... ml (10 µl) Skill required: expert No-needle techniques The term “injection” is most often associated...

  8. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    ...Importantly, conventional flow cytometry uses one detector to detect one fluorophore and therefore one cellular...fluorophores can “spill over” into detectors that are not the primary detector for that fluorophore. For example...single (primary) detector to identify a fluorophore, spectral flow cytometers use all detectors for all fluorophores...panels: five laser spectral flow machines have 64 detectors, allowing for the use of up to 64 markers in a...emitted by a fluorophore is guided to a single detector using a series of mirrors and filters. Importantly... FITC signal is spilling over into the primary detector for PE. In order to correct for this, compensation...the fluorescent molecules can be quite broad, detectors will usually only pick up a small portion of that...

  9. Getting to Know Expansion Microscopy

    Type
    Blog Post
    .... However, most SRM techniques also require specialized instruments and technical expertise, limiting ... model systems, paired with SRM techniques, and adapted for detecting nucleic acids as well as proteins...biological features previously only visible via techniques like super-resolution microscopy or electron ...Super-resolution microscopy (SRM), a family of imaging techniques developed to break through the resolution barrier...some of the limitations?  A “super-resolution” technique that uses common reagents, straightforward protocols...there is always a catch - some limitation of the technique that users should be aware of - and ExM is no ...microtubules. This issue is shared by other SRM techniques and can be alleviated by maximizing label density...

  10. How to Be an Excellent Trainee

    Type
    Blog Post
    ...learning a new technique while visiting a lab, or maybe even training on a new technique with a coworker...research group, you will also typically receive technical mentorship from a ‘bench mentor’. In some instances...mentor is a staff scientist, postdoc, lab manager, technician, or a graduate student. This is the person you...basis who will help you get on your feet with techniques in the lab (and possibly much more, depending...difficult for you to become proficient in lab techniques, which will hinder your progress. You will also...serve two purposes – familiarize you with the techniques you will be learning and the biology you will...there for you to be independent performing the technique you trained to do? Walk through the protocol mentally...

  11. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...Förster) Resonance Energy Transfer, technique which allows the detection of molecules' interactions, modifications...fluorophores to proteins, pioneers of this technique were able to directly detect protein/protein interactions in...dissociations in situ. Used since the mid-90s, this technique has revolutionised the way we apprehend molecular...playing “live.” Indeed, FRET was one of the first techniques which enabled the measurement of single molecule...microscope. Historically, molecular interactions were detected by indirect means often using probes with the ... fluorophore can be measured using microscopy techniques. FRET measurement sensitivity makes it suitable...DNA and proteins. FRET, in theory, is an easy technique, but it is very important to follow some simple...

  12. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...to get right. If you’ve thoughtfully done the technical design of your blot, but are still finding yourself...Most people will simply load an extra ladder or technical replicate (ie, leftover sample) to fill the relevant...you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to make sure you...transfer. Troubleshooting The two most common (non-technical) issues with transferring are smaller proteins...than just high background, your issue is likely technical. White circles, for example, could indicate that...incubation steps.  Many errors can arise from technical issues. The good news is that such errors are ...means the amount of protein loaded is outside the detectable range of that antibody dilution. Adjust the antibody...

  13. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ...chromosome, which was technically challenging. Other homology based technologies We've described Sequence...alternatives to restriction enzyme cloning. These newer technologies have become more and more common, and for good...Overview of Gibson assembly The Gibson assembly technique was first described by Dr. Daniel Gibson and colleagues... 5 days. One drawback to the Gibson assembly technique is that the process works best with fragments ...Gibson assembly combined with the popular CRISPR technology and is described in a recent publication by the...When followed by the standard Gibson assembly techniques described above, this resulted in the direct ...Cloning (System Biosciences) In-fusion Cloning (Clontech) Although these kits may come at a high price...

  14. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...Fluorescent imaging techniques have become indispensable tools for molecular and cell biologists over... Osaka University have developed Nano-lantern technology. Nano-lanterns contain a Renilla luciferase variant...but it has precluded widespread adoption of the technology. In 2012, Saito et al. described the first Nano-lantern...luciferase (RLuc8) with Venus. This luminescent technology is based on the principle of bioluminescence ...to excite the fused fluorescent protein. This technique is very similar to the widely used FRET, but it...ATP sensors based on Nano-lantern, showing the technique’s versatility and adaptability to many different...optogenetic system. Biosensors based on Nano-lantern technology are also very robust, with an eNL-based calcium...

  15. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...please read on to learn more about this exciting technology from the folks at Transposagen. This post was...engineering approaches. Most importantly for this technology, however, is that the transposase also enables... with no known upper limit [1]. The piggyBac™ technology can be used for numerous applications, including...of interest. Fortunately, by combing nuclease technology with the piggyBac™ system, these challenges can... DNA using the piggyBac™ transposon selection technology using a site-specific nuclease to stimulate homologous...of using piggyBac™, you could use recombinase technology (Cre/loxP) to remove the donor cassette. While...summary, the piggyBac™ transposon is a powerful technology that may be used for a variety of research applications...

  16. Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)

    Type
    Blog Post
    ...things more interesting, we are working in a new technical discipline that nobody understands (synthetic ... However, since the lab was highly focused on protecting intellectual property, it worked away in isolation...ideas (how to stop rhino horn poaching using biotechnology) were thrown on the table and dissected with...that survived the literature review, survived technical feasibility analysis, and was a marketable product... our day job, we worked on plants engineered to detect explosives and rapidly indicate their presence....reputation. The public is can be fearful of this technology and angry at the agriculture companies that produce...GMO food plants. When a press piece ran on the detector plants, we expected to see that same anger. Instead...

  17. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    ...Spectrophotometers. NEB Tech Note (2019).  Loughrey S and Matlock B. Detection of Protein in Nucleic Acid...Thermo Fisher Scientific Tech Note (2016).  Loughrey S and Matlock B. Detection of phenol in nucleic acid...Fisher Scientific Tech Note (2017).  Extinction Coefficients. Thermo Fisher Scientific Tech Tip (2013).  Additional...characteristic absorbance spectra, which you can detect by examining the A260/280 and A260/230 ratios.  ...potential contaminants, some of which are easier to detect by spectrophotometry than others. The ones most... to other reagents, making it more difficult to detect. A particularly strong absorber at 230 nm is guanidine...However, such systems are no substitute for sound techniques, routine machine maintenance, and critical interpretation...

  18. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...enhanced detection of the product. As an added benefit, replicates are built into the technology. One well...applications for the technology.  Droplet digital PCR is well suited for the detection of low copy numbers... the detection of rare sequences and single cell analysis. It is also being used in the detection of microbes...approximately 20,000 droplets using water-oil emulsion technology. Each droplet contains the ingredients for amplification... Most droplet readers have a few channels for detecting fluorescence so that it is possible to measure...applications include measuring viable probiotics and detecting circulating pathogens.  With its many applications...Central PMCID: PMC3991984. Song, Neng, et al. "Detection of circulating Mycobacterium tuberculosis-specific...

  19. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    ...molecules is necessary for a detectable signal, limiting the sensitivity of the technique. One potential solution...It seems that there’s a new CRISPR advance or technique published every week! One of the newest applications...make this system truly advantageous. This new technique, developed in Thoru Pederson’s lab, brings us ...to mapping pairs of intrachromosomal loci. The technique successfully resolved loci with physical map distances...modifications and applications Ma et al. developed this technique using standard fluorescence microscopy, and are...extending the potential imaging time for this technique. The NIH Common Fund has made mapping the 4D nucleome...of genomic loci. Ma et al. anticipate that the technique will be applicable to studies of cell cycle progression...

  20. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ...the capabilities of fluorescence-based techniques in biotechnology. Recent examples of advancements include...These dyes can also be attached to an antibody to detect various targets, such as proteins, nucleic acids..., which are pivotal in their application in biotechnology (Grimm & Lavis, 2022). These variations directly...and acceptors in their product descriptions, or technical manuals. Specific applications and matching dye...several factors. Firstly, brighter dyes enable detection of finer details by improving signal-to-noise ...designed for use in super-resolution microscopy techniques, such as Stimulated Emission Depletion (STED)...-fluorescent light when excited. While it is technically cell-permeable, its use in live cells can sometimes...
Showing: 141 - 160 of 797 results