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We narrowed to 1,006 results for: RING

Showing: 521 - 540 of 1006 results

  1. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...activity – it only requires the presence of oxygen during maturation. This meant that the protein could be...study of dynamic, physiological processes. Re-engineering GFP to increase its range of color and application...its sequence was elucidated, scientists began engineering new versions of GFP through mutagenesis in order...us how! And come back next week when we'll be featuring a guest blog post from Gal Haimovich of greenfluorescentblog...

  2. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...neuronal loop which is activated and strengthened during memory formation and memory retrieval. In neuronal...recordings. The origins of these methods are naturally occurring viruses that infect, persist, and migrate within...modified and used as tools for tracing. With some engineering, RABV and HSV, but also others including pseudorabies...plasmids at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have been widely...AAV1 – 9, with most of these serotypes capable of entering neurons (see table 1)(Choi et al., 2005; Taymans...

  3. Hot Plasmids - November 2023

    Type
    Blog Post
    ...phage-assisted continuous evolution and protein engineering to generate seven new prime editors, PE6a–g, ...(2023). Phage-assisted evolution and protein engineering yield compact, efficient prime editors. Cell,... increase the utility of this technique.  By engineering a light-sensitive LOV domain into the TurboID...mass spectrometry workflow identifies proteins bearing both labels (which had therefore been trafficked...

  4. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ... not all, of these new experimental species. Discovering ways to culture, propagate, transform, store,...systems. How the EDGE Program promotes scientific sharing The EDGE Program is designed to expand the number..., in general, these labs all include materials sharing in their dissemination plans and in particular,...to mate with and if she decides to raise their offspring or leave them to their own devices. Chytrid fungi...

  5. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...for the entire translation process. For example, during high growth rate conditions, E. coli preferentially...biosynthetic enzymes may be preferentially charged during amino acid starvation thus resulting in higher ...getting your genes of interest codon optimized by ordering plasmids containing them from Addgene. If a plasmid...that will generally segregate with the cell pellet during purification procedures. Even if you produce a ...

  6. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ... Research Trust (RSRT) has an ambitious goal of curing Rett Syndrome and Addgene is thrilled to partner...methylated CpG dinucleotides and CAC trinucleotides during neuronal maturation (Tillotson & Bird, 2019) and...neurological deficits observed in Rett Syndrome. Restoring the MECP2 gene by excision of the Stop cassette...have shown exciting and promising results for repairing mutations in models of neurological disease. Klaus...

  7. Career Insights: Technical Support Specialist

    Type
    Blog Post
    ...but did not consider them for myself. I started picturing myself in a technical support role only when I...there is a second-line team that takes on cases requiring in-depth analysis, data review, in-house testing...are obtained. Or someone may heat an ELISA plate during incubation steps in order to enhance the OD signals...with other departments such as research and manufacturing, quality assurance, product management, marketing...

  8. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...tools this could be applied to. When it comes to altering gene expression, CRISPR technologies have gradually...-RNA (miRNA) Short hairpin RNA (shRNA) Short interfering RNA (siRNA) Piwi-interacting RNA (piRNA) miRNA..._module_attribute %}{% end_module_block %}   Interfering in the future While RNAi has retreated a bit ... the background, it remains a solid option for altering the expression of specific mRNAs. shRNAs are a...

  9. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    ...biotin-streptavidin interaction (Figure 1a). This interaction brings the fluorophore in proximity to the target marker... used. These dyes are incorporated into the DNA during DNA synthesis and then diluted with subsequent ...negative live cells can still be easily gated on during the analysis. Conclusion Congratulations! You made...intracellular labeling protocol is necessary, requiring chemical treatment of the cells after surface ...

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

    Type
    Blog Post
    ... allow cells to become persistently fluorescent during development. Stochastic multicolor labeling systems... tags instead of XFP to optimize it for tissue clearing, a method that makes tissues, such as the brain...visible. Chemical tags are more resistant to tissue clearing methods and can be labeled by synthetic fluorophores...Multicolor Brainbow imaging in zebrafish." Cold Spring Harbor Protocols 2011.1 (2011): pdb-prot5546. Richardson...

  11. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...is a constantly evolving and demanding field, requiring a variety of biological and molecular tools. One... perfect for molecular cloning and metabolic engineering.  The E. coli genome has an extensive research...be analyzed to understand human disease alleles during and after embryogenesis. Addgene has over 600 plasmids...organism: quick development, a large number of offspring (produces many seeds), self fertilizing, a small...

  12. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    ... the cut site were not efficiently incorporated during HDR. We decided to systematically characterize ...References 1. Cong, Le, et al. "Multiplex genome engineering using CRISPR/Cas systems." Science 339.6121 (...Mali, Prashant, et al. "RNA-guided human genome engineering via Cas9." Science 339.6121 (2013): 823-826. ...Fidelity Cas9 Variants How to Design Your gRNA Delivering CRISPR Ribonucleoproteins Resources on Addgene.org...

  13. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...), and in SNP analysis (to introduce naturally occuring SNPs in a plasmid context). The technique is also...sequence critical for Cas9 cleavage), thereby rendering the resulting construct resistant to Cas9 induced... successfully anneal to the plasmid of interest during the PCR reaction. Ideally, your primers should ...’ll want to steer clear of these strains when preparing the plasmid template for site directed mutagenesis...

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

    Type
    Blog Post
    ...assembly work? The required homology between neighboring fragments can be created via PCR amplification...including regions of homology to the vector or neighboring fragment. Then you would amplify the fragments...required single-stranded annealing and priming of neighboring assembly fragments. Gibson assembly is commonly...modular overlap-directed assembly with linkers) which brings modularity to overlap-directed methods (Casini ...

  15. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...helpful from a biological standpoint, it makes delivering AAVs to the brain difficult. Traditionally, delivery...and/or its intermediate engineered precursor. Considering systemic capsids for your research? While systemic.../cJ and CBA/J mice. AAV1-X1 was developed by transferring the 7-mer peptide region from variable region...Fox AS. (2023). Advances in AAV technology for delivering genetically encoded cargo to the nonhuman primate...Bloom, J. M., Seed, C., & Deverman, B. E. (2019). Delivering genes across the blood-brain barrier: LY6A, a...AAV vectors to the central nervous system by engineering capsid–receptor interactions that enable crossing...

  16. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...recorded plasmid multimerization in the naturally occurring E. coli plasmid ColE1 (Goebel & Helinski, 1968...whole plasmid sequencing of our plasmid samples during our quality control process. Since MiSeq involves...D, Berg DE. pBR322-derived multicopy plasmids harboring large inserts are often dimers in Escherichia ...Guttman A, Thompson L. Vaccine Plasmid Topology Monitoring by Capillary Gel Electrophoresis. Curr Mol Med...

  17. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...An off-target rate of 5% is far too high when considering human patients and the diseases associated with...off-target frequency is (more on this below). Lowering off-target events There are several strategies...compatible with the bulkiness that accompanies mispairing of bases. To reduce off-targets, you’ll want ...which rely on insertion of DNA by the NHEJ pathway during DNA repair of the break to allow for site-specific...

  18. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    ...the five is different. To identify significant pairings, additional tests, referred to as post-hoc analysis...temperature, humidity, sunlight exposure time, and watering volume on total crop production (though you would... error is the probability of a false positive occurring when many comparisons are performed. For example...drawbacks to these tests as they can be quite stringent if many samples are assessed. However, if many...

  19. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...questions in your own lab. 2. Seek multiple mentors during your postdoc At the end of grad school, you searched...own experience, great mentors often can be found during conversations with senior investigators. Discussions...fundamental conflict. Science is about exploration, discovering new information. There are literally infinite...well practiced or too skilled at presenting and answering questions. Organizing a thoughtful engaging talk...

  20. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...minimize recombination that are typically employed during viral plasmid amplification (Fischer, KB. 2019)... showing the mechanism of spontaneous reversion during AAV plasmid amplification. On the left homologous...  Some classes of neurons transiently express TH during development or have low levels of TH mRNA without...time, frustration, and funds in the long-run by ensuring you are minimizing any off-target expression that...
Showing: 521 - 540 of 1006 results