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We narrowed to 882 results for: BASE;

Showing: 221 - 240 of 882 results
  1. How to Design Your gRNA for CRISPR Genome Editing

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    Blog Post
    ... you are trying to do: gene knockout, a specific base edit, or modulation of gene expression. Location...with BioRender.com. The jigsaw: Editing by HDR, base editing, and prime editing For a specific edit, ...constraints are even more exquisite for the so-called base editor Cas9, which makes DNA changes in the absence...dsDNA breaks (Rees et al., 2018). For C>T and A>G base editors, the intended edit must be in a 5 - 10 nt...others have examined the ability to use sequence-based and other features to nominate gRNAs that are likely...(2020) Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors. Nat Biotechnol...doi.org/10.1093/nar/gkw583 Rees HA, Liu DR (2018) Base editing: precision chemistry on the genome and transcriptome...
  2. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...optogenetics, such as channelrhodopsins and tools based on light-oxygen-voltage (LOV) domain. They can also...interactions, we recommend using NIR split reporters based on miRFPs (Shcherbakova et al., 2016), since they...Filonov et al., 2013). FRET biosensors were developed based on miRFP760-miRFP720 FRET pair, including Rac1 biosensor...intensiometric calcium indicators NIR-GECOs were developed based on mIFP6. Learn more about the characteristics, ...Brightness in HeLa cells vs. iRFP713, % d Ref BphP-based FPs miRFP670 https://www.addgene.org/79987/ 642...720 98,000 6.1 97 Monomer 510 4.5 117 12 CBCR-based FPs miRFP670nano https://www.addgene.org/127443...
  3. Finding nucleic acids with SHERLOCK and DETECTR

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    Blog Post
    ...has been started based on DETECTR technology with the mission to “provide a CRISPR-based platform on which...SHERLOCK and DETECTR demonstrate the power of CRISPR-based diagnostics. SHERLOCK- Specific High Sensitivity... DNA or RNA with atomolar sensitivity and single base pair mismatch specificity.  SHERLOCKv2 SHERLOCK ...isothermal amplification by RPA. When Cas12a-cRNA base pairs with the dsDNA of interest, the DNase activity...Wadford DA, Chen JS, Chiu CY (2020) CRISPR–Cas12-based detection of SARS-CoV-2. Nature Biotechnology. https...
  4. Antibodies 101: Introduction to Gating in Flow Cytometry

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    Blog Post
    ...different parameters that your gating strategy will be based upon. We’ll try to keep it short though, promise...SSC: scattering To digitally extract your cells based on their size, the cytometer provides you with two...lymphocyte population (10.7% of all recorded events) based on their expected size. The strong signals close...not of interest can still be gated out later. Based on personal preference, some people like to make...staining signals. For this, you’ll need to gate based on the proportion between H and A (or W and A) within...When you’re interested in measuring a population based on a fluorescent stain, you might be wondering which...
  5. Luminescent Imaging with Nano-lanterns

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    Blog Post
    ...system. Biosensors based on Nano-lantern technology are also very robust, with an eNL-based calcium sensor...RLuc8) with Venus. This luminescent technology is based on the principle of bioluminescence resonance energy...They also developed calcium, cAMP, and ATP sensors based on Nano-lantern, showing the technique’s versatility...
  6. Michael Koeris' Journey from Grad Student to Entrepreneur: The Story of Sample 6

    Type
    Blog Post
    ...time was not right for the development of phage-based therapeutics. After a brief stint at the venture...from developing therapeutics to developing phage-based tools for detecting pathogens in food products. ...biotechstart.org Read Sample 6's Paper on Bacteriophage-based Diagnostics Additional Resources on the Addgene...
  7. Open Enzyme Collection

    Type
    Collection
    ...harder to obtain. The enzymes can also be used as a base to improve functionality or add new functions through...
  8. Viral Production

    Type
    Collection
    ...determined using a chromogenic endotoxin detection assay based on the amebocyte lysate method. Purity Purity of...
  9. CRISPR in the Clinic

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    Blog Post
    ...initiating a base editor-based clinical trial for SCD in 2022 and Editas Medicine starting a Cas12-based clinical... today. Below we will review some of the CRISPR-based editing therapies that are currently approved or...
  10. DNA Purification Without a Kit

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    Blog Post
    ...gravity-based column plasmid maxiprep kit takes 1-1.5 hours to complete, while this syringe-based method...Compared to a commercial magnetic bead-based kit, this cellulose-based method was faster, cheaper, and had... Addgene follows a similar workflow as a column-based plasmid extraction kit. First, you lyse the bacteria...
  11. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

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    Blog Post
    ...are many protein-based toxin-antitoxin systems. One of the most well known protein-based toxin-antitoxin...counter this, methyltransferases add methyl groups to bases at specific nucleotide sequences, preventing the...system that is regulated by antisense RNA, which base pairs to cognate mRNA to inhibit its translation...Crystal structure of the MazE/MazF complex: molecular bases of antidote-toxin recognition." Molecular cell 11.4...
  12. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ... let the virions out. We recommend using a Tris-based lysis buffer and sonication.  The obtained virions...particles that can transduce cells, which can vary based on target cells and can be altered by freeze/thaw...experimental outcome and planning.  Both ddPCR- and qPCR-based titering assays measure physical titers, meaning...post.  We generally do not recommend using ELISA-based titering assays, since those will quantify both ...Inactivation of Adeno-Associated Viral Vectors by Oxidant-Based Disinfectants. Human Gene Therapy, 32(13–14), Article...
  13. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ...for studying colloidal gold (1).  The method was based on using a thin plane (sheet) of light generated...microscopes, optical sectioning of a specimen is based on discriminating the out-of-focus reflected light... combining Ultramicroscopy with tetrahydrofuran-based tissue clearing (13,14). A few years later, Karl...Deisseroth’s lab developed CLARITY, which is a hydrogel-based method that also allows antibody penetration for...recently developed AAV (adeno-associated virus)-based fluorescent labeling methods that can be used with...
  14. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...desktop-based software for plasmid design and mapping. ApE (A plasmid Editor): A free, donation-based plasmid...knowledge resource for cell lines IBEX Knowledge Base: A data repository for validation data on IBEX reagents...antibodies: A database of validated antibodies for mouse, human, and marmoset brain IHC.  FPbase: A data repository... Flame plasmid pET21a-BirA.   DNA / protein database search and analysis tools Addgene's analyze sequence...sites Open reading frame finder  Addgene vector database: Find sequences for plasmids assembled from publications...nucleotide or protein sequences to sequences in a database ExPASy translation tool: Translates a DNA sequence...sequence using all six open reading frames UniProt: A database of protein sequence and functional information...
  15. 8 Great Depositor Research Stories

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    Blog Post
    ...tells us about a new tool for efficient multiplex base editing and multiplex prime editing (requiring minimal...dCas9 (with no modifications!) as a CRISPR/dCas9-based tool for DNA demethylation. AAV Q&A with Tim Miles...
  16. Plasmid Tools for Microbiome Studies

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    Blog Post
    ...microbiome studies.  Find the MAGIC plasmids here Paper-based synthetic biology approach to analyze gut microbiota...microbiome, James Collins’s lab developed a paper-based method that can be used to detect microbes and biomarkers...As mRNA expression cannot be identified with DNA-based qPCR methods, the ability to detect mRNA transcripts...Takahashi, Melissa K., et al. "A low-cost paper-based synthetic biology platform for analyzing gut microbiota...
  17. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...slightly with either a 1-7 base poly(U)tail in tRNA-gRNA constructs or a 1-4 base tail from the second tRNA... cassettes (considering the gRNA itself is ~100 bases) limit the number of gRNAs that can be packaged ...Pseudomonas aeruginosa to cleave 4 gRNAs separated by 28 base Csy4 recognition sites from a polycistronic transcript...
  18. Plasmids 101: Cre-lox

    Type
    Blog Post
    ...LoxP (locus of X(cross)-over in P1) sites are 34-base-pair long recognition sequences consisting of two...event. This event can have three general outcomes based on the location and orientation of the loxP sites...above, a genetic modification can be restricted based on experimental constraints. This has been used ...asymmetric spacer "NNNTANNN", where "N" indicates which bases may vary from the canonical sequence. Among these...
  19. Technical Design of a Western Blot

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    Blog Post
    ...options come in two categories: protein-based and chemical-based. Before selecting one, check your antibody...concentration. That makes the choice easy! Protein-based blockers include dry non-fat milk or serums like... of blocker, you can also use gelatin.  Protein-based blockers are inexpensive and easily made in the ... tip! If you get a speckled image, your protein-based blocker is either degraded or not completely mixed...mixed together.   Commercially available chemical-based blockers, while much more expensive, are consistent...to another. Since antibody binding varies wildly based on proteins, and their differing amounts of available...Smaller dynamic range; no multiplexing Protein-based blocker Degrades quickly Wet transfer Increased...
  20. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...short ncRNAs, such as microRNAs that are around 20 base pairs long. Using a single guide RNA (sgRNA) to ...Figure 1: A schematic illustration of CRISPR/Cas9-based approaches to study ncRNAs. (A) Transcription start...your experiments, there are plenty of other CRISPR-based options that can target ncRNAs.  CRISPRi and CRISPRa...10.1126/science.1225829  Hazan J, Bester AC. CRISPR-Based Approaches for the High-Throughput Characterization...
Showing: 221 - 240 of 882 results