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We narrowed to 236 results for: SPL

Showing: 21 - 40 of 236 results

  1. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...contaminants. Set up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine..., for example, split cells on a Monday diluting them 1:10, you should be able to split on the following...Thursday, or, at the latest on Friday. It’s best to split cells before the plates are more than 80-85% confluent...metabolism that is toxic to the cells), and need to be split very soon. There is power in numbers! When starting...

  2. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...reporters altogether; In the split-luciferase system, intein-mediated splicing reconstitutes functional luciferase... luciferase protein interact and enable intein splicing (5). Screening with yeast two hybrid systems While... this requirement for transmembrane proteins, a split ubiquitin system has been devised (8). References...Kaihara, A., Sato, M., Tachihara, K., Umezawa, Y.l. Split luciferase as an optical probe for detecting protein-protein...interactions in mammalian cells based on protein splicing. Anal Chem. 2001; 73:2516-2521. Pubmed PMID: 11403293...

  3. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    ...reporter, ZipGFP, which utilized the self-assembling split GFP (GFP1-10 and GFP11) to increase fluorescence... this they modified a different self-assembling split GFP (GFP1-9 and GFP10-11) by flipping a single β-strand...11 β-strands and one central ɑ-helix that can be split into 3 parts: (1) β-strands 1-9 (β1-9) and the one...superfolder Cherry (sfCherry). Like FlipGFP, sfCherry was split into  β1-9 and β10-11, where β10-11 was forced into...-protein interaction sensor based on tripartite split-GFP association." Scientific reports 3 (2013): 2854...

  4. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...excited to announce that our plasmid and sequence displays are now powered by GSL Biotech's SnapGene Server...feature library, our updated plasmid and sequence displays are now much easier to interpret and analyze at...CRISPR gRNA cloning. With this more functional display, you can start thinking about your next cloning...experiment early on. Beyond these simple but powerful display improvements, when you click on any plasmid map...enhancements that will further improve the map displays and functions. ...

  5. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ... some you’ve likely never heard of. Each entry displays the following information for a Cas enzyme: ...properties Links to relevant resources The homepage displays a graphic describing CasIDs and relevant properties...feature. Activity features: CasID CasPEDIA will display both the cis and trans activity of each entry. ...protospacer-flanking sequence (PFS) in some cases. The resource displays these requirements in regard to their sequence... covered!     Fig. 1 – CasPEDIA Cas ID display information for a given Cas (SpyCas9a) from CasPEDIA...

  6. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...scientists can mutate the transcription start site, splice sites, exons, and/or promoters to knock out lncRNA... and Bester 2021; Zibitt et al. 2021). Mutating splice sites is particularly effective as it can result...Transcription start site (TSS) deletion. (B) Mutations of splice sites. (C) Removal of an exon or a large genomic...noncoding RNAs in human cells by Cas9 targeting of splice sites. Nat Biotechnol. Published online November...expression by lncRNA promoters, transcription, and splicing. Nature. 2016;539(7629):452. doi:10.1038/nature20149...

  7. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    Blog Post
    .... This style displays a poster vertically rather than horizontally. Instead of displaying your information...employing a large central box, often used for displaying all of one’s figures, or at least the majority...inserted. The advantage of this style is that it will display your data most prominently, making it the star ...information as columns, you may choose to display it as rows. It can be more catchy because it is generally...

  9. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...range of PAMs like NG, GAA, and GAT, but also displaying increased editing specificity. We’re excited ...recognizing NG, NNG, GAT, and CAA PAMs, while xCas9 3.6 displayed the second best PAM range. Characterizing xCas9... SpCas9 in mammalian cells, finding xCas9 3.7 displayed about the same editing rate as SpCas9 at an NGG...SpCas9. For some non-promiscuous gRNAs, xCas9 3.7 displayed more than a 100 fold decrease in off-target editing...ratio. Importantly, the observed off-target loci displayed a broad range of PAM sequences, indicating they...

  10. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...detecting protein-protein interactions that uses split fluorescent proteins tethered to potential protein...visualize PPIs. This fluorescent protein (FP) is split into two fragments that individually lack fluorescence...fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent protein, YFP) is reunited...information about the structures of the tested PIPs. Many split FPs have been validated for BiFC (Kodama and Hu,... Hu, 2012). Such controls are necessary because split FPs can associate, albeit at relatively low frequencies...

  11. Antibody Plasmid Collection

    Type
    Collection
    ... 2018;1827:313-334. Daniel Christ pComb3 phage display vector system for producing human antibodies Synthetic...1995). Carlos Barbas Variant of the pComb3 phage display vector system for isolation of scFv, diabody and...chicken monoclonal antibody fragments by phage display. J Immunol Methods. 2000 Aug 28;242(1-2):159-81...

  12. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...anchoring domains The original iGluSnFR in the pMinDisplay backbone is fused to a specific peptide segment...TMD) that anchors the sensor to the membrane and displays it on the extracellular side. In addition to the... 4 Sequence schematics for PDGFR, GPI, and SGZ display constructs. Image adapted from Aggarwal et al… ...transmembrane domain in the mammalian expression pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol...

  13. Cre-Lox and Other Site-Specific Recombinases

    Type
    Collection
    ...drug-inducible approach may also be used. Split proteins: The recombinase is split into inactive N- and C-terminal...or search by inducible system (e.g., search for "split", "rapamycin", “tamoxifen”, or “light”): Cre Recombinase...

  14. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...array targeting the MAPT exon 10 splice acceptor and two putative splice enhancers. After AAV-mediated delivery...previously characterized Cas13 enzymes, CasRx also displays very high on-target efficiency; where shRNA treatment...produced 500-900 significant off-targets, CasRx displayed zero. Unlike Cas9, for which efficiency varies...

  15. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...following a G more efficiently than BE3, but APOBEC3G displayed less predictable sequence preferences.  The Liu...editors (ABEs). ABE7.10 is the most active editor, displaying an average editing efficiency of 53% with an ...of target positions 4-7. ABEs 6.3, 7.8, and 7.9 display slightly wider editing windows of position 4-9,...produce a mixed population of edits, ABEs do not display significant A to non-G conversion at target loci...Highlights Komor et al., 2016 BE1, BE2, BE3   BE3 displays highest editing efficiency but higher indel formation...

  16. Pooled Library Amplifications

    Type
    Blog Post
    ...from the Mark Dawson lab Re-transformation of the SPLINTR Libraries Protocol.   Selecting your strain...perform several individual transformations, such as splitting 400 ng of DNA in 100 μL of cells over four individual...Electroporation Protocol (DOCX, 15.8 KB) Barcode SPLINTR Pooled Libraries 200,000–1,200,000 60 bp ... 200 ng Liquid Re-transformation of the SPLINTR Libraries (PDF, 401 KB) Screening Zhang Lab...

  17. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...can be found at Addgene. Split FPs – some FPs (e.g. GFP, Venus) can be split into two halves, which are...proximity, they will form the full FP and fluoresce. Split FPs can be used to determine the proximity of two...two proteins fused to the halves of the split FP. This technique is also is also called Bimolecular Fluorescence...

  18. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...fusion proteins for imaging, tracking, modulating splicing, and regulating expression of specifically targeted...described in bacterial cells, LwaCas13a does not display nonspecific RNA cleavage activity in eukaryotic... LwaCas13a to shRNA, they found that LwaCas13a displays similar knockdown efficiency but no significant...to see this group of enzymes continue to make a splash in the CRISPR field. We feel that the culture of...

  19. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...cleavage frequency. Truncated gRNAs: Truncated gRNAs display less off-target activity. High fidelity Cas9s: ...Cas9 variants , SpCas9-HF, eSpCas9, and HypaCas9 display very low off-target activity due to rationally ...without creating a double-strand break. This method displays higher efficiency than HDR-mediated editing. The...

  20. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ... of interest do not lie beyond the dimensions displayed in the plot. (B) A more restrictive gate results... second measurement to choose for graphically displaying your events. You are free to choose any size-... the positioning and shape of the populations displayed. As the separation pattern largely stays the same... cells but not dendritic cells. Using a graph displaying both stains, I can quickly separate T killer ...
Showing: 21 - 40 of 236 results