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We narrowed to 479 results for: multi

Showing: 61 - 80 of 479 results

  2. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...single-component nuclease to mediate cleavage rather than the multi-subunit complex employed by class 1 systems. A putative...vectors. In 2017, Zetsche et al. developed a Cpf1 multiplexing approach using a single crRNA array to express...varies widely with gRNA sequence. Both groups used multiple methods to examine Cpf1 off-target editing. First...publication at Addgene. 2. Zetsche, Bernd, et al. "Multiplex Gene Editing by CRISPR–Cpf1 Using a Single crRNA...

  3. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    Published
    July 16, 2015, 2:30 p.m.
    ...adjusting the thickness of the laser sheet and using multi-view imaging can provide enhanced resolution (8,...combined light sheet microscopy with sequential multiple-view reconstruction (5). In order to image large...light scattering and absorption.  Finally, the photomultipliers used to detect light in confocal microscopes...acquisition is performed using cameras with electron multiplying couple-charged device (EMCCD) or complementary...through Addgene.  The combination of LSFM with multiple tissue clearing strategies and neuronal labeling...

  4. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...savings, and 3) ability to perform single site or multi-site saturation mutagenesis with the same protocol...mutational scanning to simultaneously test how multiple amino acid changes affect a protein of interest...a selection system. You can test the effect of multiple mutations in combination and possibly discover...that tile across a region of interest, generating multiple mutants in one PCR reaction. Below is a step-by-step...randomized bases (NNN) at a given codon site and multiple primer sets tile across the protein or region ...where it does. 1) When the backbone vector has multiple BbvCI sites. If these sites have opposite orientations...

  5. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...containing the sgRNA target sequence can be placed in multi-cloning site (MCS) between EGFP fragments. The pX330...-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering...

  6. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging. Reports... Nano-Lanterns  Suzuki, K., et al. (5) imaging multiple proteins at the same time in live cells using ..., to produce a series of FP-RLuc8 fusions for multicolor imaging. In all of these fusions coelenterazine...the Enhanced Nano-Lanterns including monitoring multiple cellular events such as the dynamics of subcellular... of bright luminescent protein for real-time multicolour bioimaging. Nature Comm. 7, 13718. PubMed PMID...

  8. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    Published
    March 14, 2017, 2:30 p.m.
    ...immunotherapeutic target is never the answer to the multi-level barricades put up by cancer. As mentioned ... use the blood stream to commute to and invade multiple organs, and most importantly escape the immune...therapies must be explored. When attacking cancer, multiple immunotherapy targets can be utilized at the same...

  9. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...Protein Imaging Choosing Your Fluorescent Protein for Multi-Color Imaging Catch Up on Your GFP Background Resources...state (“quenching”). The photoswitch can occur multiple times by activating with two distinct wavelengths...experiment even begins! If you are performing multicolor experiments, you will want to be sure that the...relies heavily on the same fluorophore being imaged multiple times in order to reconstruct an image. The best...Photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy...Collection Find Photoactivatable FPs for Single and Multicolor in vivo Imaging Browse All Fluorescent Protein...

  10. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    ... indels Cytosine transversion editors rely on a multi-step cellular repair pathway that is known for sometimes...that site is filled depends on cellular factors. Multiple labs tried to identify beneficial factors to include...Several of these proteins had additive effects when multiple were included in the CGBE. They also developed...

  11. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...indicated Synaptotagmin-1 antibodies and corresponding Multi-rAb CoraLite® Plus 647 secondary antibodies. Antibody...antibody validation, and it is recommended to screen multiple candidate antibodies when possible. If cost is...techniques — and so it may be necessary to try multiple techniques. For example, we have found that some...potentially the need to study protein expression at multiple timepoints. When the target of interest is critical...challenging. Correlation with RNA or Proteomic data from multiple cell lines/ types with different expression Flow...with an assessment of labeling pattern between multiple antibody clones targeting the same protein (independent...be particularly useful when designing larger multicolour panels, where different manufacturers may have...

  12. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ...donor DNA. By using a multi-spacer CRISPR array, CRISPR-transposons can be multiplexed. Transcriptional repression...is a fast, multiplexable method that combines CRISPR and recombineering-based MAGE (Multiplex Automated...functions in S. pneumoniae and can be used to generate multiple mutations simultaneously (Jiang et al. 2013). ...mutations and 66% efficiency for small insertions. Multiplexing of two targets simultaneously is possible with...are its regulatable nature and its ability to multiplex repression. Using a Tet responsive promoter to...cassette, one can reliably knockdown expression of multiple target transcripts simultaneously. Marraffini ... system results in 80-90% RNA knockdown across multiple gRNAs per gene (Singh et al. 2016). Sarah Fortune's...

  13. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...improve the ease and convenience of assembling these multi-protein fusions or “polyproteams”, the Howarth lab...stimulate neurons. Recent studies have designed multi-component actuators that are sensitive to radio ...it comes to simultaneously modifying multiple loci (multiplexing). Currently, the only genome engineering... and basic biologists alike often need to test multiple expression systems before they achieve the appropriate...have greatly simplified the process of generating multiple E. coli gene expression plasmids with the Cross-disciplinary...library design, Moffat laboratory members conducted multiple screens in different human cell models and identified...method for bacterial genome engineering across multiple species Recent discoveries in the fields of genome...

  14. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    Published
    Aug. 10, 2021, 1:15 p.m.
    ...Quantification.”   Multiplex PCR Multiplex PCR, as the name implies, is a method in which multiple targets can...forward and reverse primer sets.  Multiple template PCR - multiple templates with different primer pairs...Scientists can also utilize multiplex qPCR to quantify the concentrations of multiple starting DNA templates... changes in target DNA levels Multiplex PCR Amplify multiple targets from one sample Disease or...practice in molecular biology and can be used in a multitude of scientific techniques such as molecular cloning... be amplified in a single PCR experiment using multiple primers all in one PCR reaction. This is an extremely...laboratory.  There are two main categories of multiplex PCR:  Single template PCR reaction - one template...

  15. Deep Dive: qPCR

    Type
    Blog Post
    Published
    Sept. 29, 2022, 1:15 p.m.
    ... If there are multiple amplicons, or primer dimers forming, the curve will show multiple peaks.    ...optimizing for LightCycler machines.) Multiplexing It is possible to multiplex qPCR when using fluorescent probe-based...change, the multiplexed protocol may have to be re-optimized.  While troublesome, multiplexing your reference...the Lightcycler while being relatively easy to multiplex (Thellwell, 2000). See Table 1 for a comparison...noise Probe Allelic specificity Can be multiplexed Single amplicon validation technique Hydrolysis... reagents in your chemistry, unless running a multiplex reaction or on a rapid cycle machine. Melt Curves...data will be much improved if you move to using multiple reference genes whenever possible (Vandesompele...

  16. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    Published
    April 7, 2015, 4:21 p.m.
    ...transcriptional programs on multiple genes. Modifying the CRISPRi system for multidirectional control The CRISPR...coli. The utility of simultaneous and multidirectional multiplex control via scRNA design is readily apparent...targeted site. Multiple scaffold RNAs can be designed to allow simultaneous, multidirectional and tunable...discussed in the paper. The transcriptional control of multiple loci is deftly coordinated by the eukaryotic cell...toward tinkering with regulatory networks and multiplex gene control. However, these approaches are often...Modifying the CRISPRi system for more flexible multidirectional control, a new study repurposes the sgRNA ...scaffold modules are comparatively small allowing multiple sgRNA constructs to be introduced simultaneously...

  17. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ...visualize multiple targets at once. An advantage of multiplexing is that you can ask how multiple proteins...and colors makes it easy to perform multiplexed IF. By using multiple primary antibodies made in different...antibodies are generally polyclonal, which means that multiple secondary molecules can bind to a single primary...species and to label targets with different colors. Multiplex immunofluorescence Speaking of different colors... excitation/emission spectra do not overlap. Multiplexed IF can also require some additional optimization...antibodies. But, once you do find the right protocol, multiplex IF can give you impactful and beautiful results...minimal impact to the target epitope. There are multiple methods for fixing samples, each with its own ...

  18. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    Published
    Oct. 5, 2021, 1:15 p.m.
    ...for Biological Studies) Multiple uses: From multiplexing to multiscaling NIR-shifted spectrum is perfect...perfect for use in spectral multiplexing (Shcherbakova et al., 2018). NIR FPs can be combined with blue-light...These probes can be combined in two-color and multicolor NIR imaging.  Small miRFPnano A separate class...Verkhusha, V.V. Near-Infrared Fluorescent Proteins: Multiplexing and Optogenetics across Scales. Trends Biotechnol...fluorescent proteins as tags and biosensors for multiscale imaging. Nat Commun 7, 12405 (2016). Qian, Y....monomeric near-infrared fluorescent proteins for multicolor imaging across scales. Nat Commun 11, 239 (2020...cyanobacteriochrome as versatile tag for spectral multiplexing. Nat Commun 10, 279 (2019). Oliinyk, O.S. et...

  19. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...Combining multiple modifications A key feature of IVA cloning is the ability to combine multiple modifications...vivo to produce the multiply modified product. For example, by combining multiple primers, a C-terminal...GFP-tag (Figure 3). Achieving multiple modifications requires multiple recombination events to assemble...regions can be encoded in the primer sequences. Multiple plasmids can be included in a single PCR, for ...of the strain: when asking the cell to perform multiple recombination events, all linear fragments need... 107 CFU/μg are perfectly fine, yet performing multiple modifications will require ultracompetent (109..., and point mutagenesis to complex assembly of multiple genes. Primer design for each type of modification...

  20. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    Published
    April 13, 2021, 1:15 p.m.
    ...for multicopy chromosomal integration (ex: for protein expression) (Zhang et al., 2020). Multicopy chromosomal... Most recently, we’ve developed a new system “Multicopy chromosomal integration using CRISPR-associated...including concatenations into crRNA arrays targeting multiple different sites in E. coli. pCutamp cures the ...Ptet, and Para. The pDonor plasmid pRE57-Ter with multiple terminators can serve to interrupt genes (or gene.... Figure 2: (A) MUCICAT plasmids for multicopy integration. (B) Anhydrotetracycline (aTc) induces...following advantages: Fast and site-specific multicopy chromosomal integration of cargo without using...dosages to facilitate swift optimization of strains. Multiple induction systems to choose from. Ready for metabolic...
Showing: 61 - 80 of 479 results