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We narrowed to 587 results for: des.1

Showing: 281 - 300 of 587 results

  1. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    Published
    Oct. 1, 2015, 2:30 p.m.
    ...animal will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models... can be explained primarily in two different ways 1) your hypothesis is wrong or 2) there is some problem...follow him on Twitter @derekpsimonphd.    References 1. Stuber, Garret D., et al. "Excitatory transmission...other words, your optogenetic experiment should be designed around a behavioral test in which your circuit...of mouse transgenics. Everything that has been described is intended for the mouse; however, all the steps....Diester, Ilka, et al. "An optogenetic toolbox designed for primates." Nature neuroscience 14.3 (2011)...

  2. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...the tag sequence flanked by homology arms (Figure 1). In the first step of optimization, we started off...form the ribonucleoprotein complex (csRNP, Figure 1). We use in-house purified Cas9 protein, which, in...standard immunocytochemistry (ICC) assays.  Figure 1: Three ingredients of epitope-tagging experiments....daydreaming about sunny days in the UK.   References 1. Bressan RB, Dewari PS, Kalantzaki M, Gangoso E, Matjusaitis...knock-in. Another good design tool can be found at Desktop Genetics .For ssODN design, remember to incorporate... systems, you will have to design your crRNAs using other web-based design tools (for C-terminus tagging... not work in pull-down assays: Epitope tagging provides a solution Proteins - the workhorses of the cell...

  3. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    Published
    July 7, 2015, 3:36 p.m.
    ...sequences are inserted into the genome. Figure 1: Self-excising cassette schematic from Dickinson et...):1028-34. doi: 10.1038/nmeth.2641. Epub 2013 Sep 1. PubMed. Find plasmids from this publication at ... the summer of 2013, a remarkable nine papers describing CRISPR/Cas9 genome engineering methods for C....America. To use this SEC-based system, first design and clone 500-700 bp homology arms for your gene...injected 60-80 animals per gene, they obtained the desired insertions on the first attempt for 6/7 genes. ...

  4. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    Published
    Nov. 3, 2015, 3:30 p.m.
    ... this blog post. I separate sgRNA design tools into database and de novo solutions. Database tools allow...automated rational design. The first of only three such databases, EENdb (published 1 Jan 2013, no longer...community in order to succeed. With respect to de novo sgRNA design, I again separate it into two categories...follow him on Twitter @CMacPhD.          References  1. Ghorbal, Mehdi, et al. "Genome editing in the human...single-target design”, “multi-target design”, “off-target aware”, “high mismatch limit”, “approximate design”, “... “empirical design”, “single-PAM design”, and “multi-PAM design”. Trends: By the end of 2014, tools began...with the post-design process such as primer and plasmid design. Examples:  “multiplex design”, “multi-method...

  5. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    Published
    March 31, 2020, 1:15 p.m.
    ... RNAs inhibit plasmid replication.   Figure 1: The replication problem. Iterons are repeated ...actually does, and neither plasmid is maintained (Figure 1).  The partitioning problem Plasmid incompatibility...Agrobacterium tumefaciens. Genes & Genetic Systems 77:1–9 . https://doi.org/10.1266/ggs.77.1 Pubmed PMID: ... Environmental Plasmids blog). Scientists have designed small, high copy incompatible plasmids that lead...Pubmed PMID: 25889267 Ebersbach G, Sherratt DJ, Gerdes K (2005) Partition-associated incompatibility caused...

  6. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ...Check out Addgene's New Viral Service! References 1. Hay, R.J., Macy, M.L. & Chen, T.R. “Mycoplasma infection...contaminations.” Methods in Molecular Biology 946 (2013). 1-13. Pubmed PMID: 23179822. 5. Segura, M.M., Kamen,...Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009...best to start from scratch. Obtain a vial of the desired cell line from a reputable distributor such as ...extreme changes in osmotic pressure and pH can destabilize the viral envelope. One drawback of the multiple...

  7. New Norepinephrine Indicators: nLightG and nLightR

    Type
    Blog Post
    Published
    Dec. 12, 2023, 2:15 p.m.
    ...nLightR (Kagiampaki et al., 2023).     Figure 1: Properties of nLightG and nLightR. A, Structural ...Article 6. https://doi.org/10.1038/s41592-022-01488-1 More resources on the Addgene blog Fluorescent proteins...will need to be accounted for in your experimental design. Find nLight plasmids here! Next steps While nLightG...

  9. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    Published
    May 21, 2020, 1:15 p.m.
    ...Rodríguez et al., 2011).  Genetics of SPARC Figure 1: SPARC and SPARC2 use the GAL4-UAS system along with... several common genetic tools in Drosophila (Fig. 1):  GAL4-UAS, a transcription factor binding site....References: del Valle Rodríguez A, Didiano D, Desplan C (2011) Power tools for gene expression and clonal...

  10. CRISPR Guide Update

    Type
    Blog Post
    Published
    Oct. 10, 2024, 1:15 p.m.
    ...refreshed figures made using BioRender.com. Figure 1: PRIME editing Please check out the updated guide...update, we have added new sections to share and describe some incredible new CRISPR technologies developed...much-anticipated prime editing section and additions describing CRISPR’s ability to make large genome edits using...transposases or the Cascade-Cas3 system. We also describe newly engineered Cas9s with more flexible requirements...

  11. Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC

    Type
    Blog Post
    Published
    Sept. 21, 2023, 1:15 p.m.
    ...following three strategies might help!     Figure 1: Species on species staining can be a problem because...same host species as your secondary antibody since 1) your secondary should not recognize Fab fragments...immunocytochemistry (ICC). Not all of the solutions described here may be as effective for other techniques,...

  12. Summer SciComm: Science Storytelling

    Type
    Blog Post
    Published
    Sept. 16, 2022, 1:15 p.m.
    ...defined as (1) the struggle between two opposing forces or (2) the result of competing desires or (3) the...journey even when they know the ending.    Fig. 1: We all know that the superheroes win in the end. ...

  13. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    Published
    Aug. 11, 2020, 1:15 p.m.
    ...and take a look at each of its parts. Figure 1: Parts of an AAV transfer plasmid. Parts shown in ..., ubiquitous expression, while the human synapsin 1 promoter is neuronal specific (Haery et al., 2019)...this post, you’ve got the knowledge to be able to design a construct for the successful and specific expression...

  14. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    Published
    Nov. 8, 2018, 3:47 p.m.
    ...from distantly related organisms. In 2006, less than 1% of the genes deposited at Addgene were synthetic....synthetic biology is expanding rapidly.  References 1. Kunjapur, Aditya M., Philipp Pfingstag, and Neil ...of 173 known synthetic and natural genes.  In designing and optimizing their algorithm, they found that...

  15. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    Published
    Aug. 8, 2019, 1:19 p.m.
    ...the journal-specific guidelines.  Peer-review step 1: The first read At the first read of the paper, I ...group of Prof. Mark Tester, where she focuses on (1) salt-induced changes in root-to-shoot ratio in Arabidopsis...of my review - "The manuscript by ZZZ et al., describes ... The authors share a wealth of data on ... ...Additionally, you can ask that data and resources described in the manuscript are accessible to the community...

  16. Evolution of Lab Techniques

    Type
    Blog Post
    Published
    June 21, 2016, 2:30 p.m.
    ...blotting, named for its inventor Edward M. Southern (1), can be used to detect DNA identity, size, and abundance...hours at low cost with restriction mapping (Figure 1). Though some labs may prefer to send DNA directly... serotonergic system and behavior.     References 1. Southern, Edwin Mellor. "Detection of specific sequences...the order of nucleotides in a string of DNA (7). Sanger sequencing uses modified nucleotides, called dideoxynucleotides...the widespread use of Sanger sequencing. Building upon this technology, radiolabelled nucleotides were...causes sickle cell anemia. The mutation happens to destroy a sequence recognized by the restriction enzyme...synthesis reaction along with all four regular nucleotides results in the synthesis of DNA strands of all...

  17. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    Published
    Feb. 21, 2019, 1:15 p.m.
    ...native endonuclease activity degrades the displaced strand.   Figure 1: EvolvR consists of an error-prone...mutation rate and extend the editing window of EvolvR: (1) two additional mutations to PolI3M generated PolI5M...diversification of all nucleotides at a user-defined loci in bacteria. Its modular nature provides versatility in...provide a means to continuously diversify all nucleotides within a user-defined genomic region. EvolvR,...method developed by the Dueber Lab at Berkeley, provides a new approach for generating novel genetic variants...users to achieve targeted diversification of all nucleotides within a defined region of a genome. Require...turbidostat. Only mutate particular types of nucleotides. Both the Bassik Lab and the Chang Lab have created...

  18. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    Published
    Sept. 28, 2023, 1:15 p.m.
    ...overcomes many of those limitations (Figure 1). Figure 1: Opto-katanin induces fast, localized microtubule...GTPase Signaling Toolbox. ACS Synthetic Biology, 11(1), 515–521. https://doi.org/10.1021/acssynbio.1c00604...optogenetics than neuronal activity. Researchers have designed a variety of photoswitchable elements into many... and limitations – for example, microtubule-destabilizing drugs are impossible to target to specific subcellular...plasmids if you want to make a stable cell line. This design allows highly localized, rapid and reversible microtubule...Rho GTPase activity. A) Schematic of Opto-RhoGEF design and activity. B) OptoTIAM (magenta) expressed in...processes in cells and tissues. And, this Opto-RhoGEF design could be extended to other GTPases. The Chow Lab...

  19. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    Published
    April 9, 2019, 1:13 p.m.
    ...sticks out and can be considered a linker (figure 1). In fact, for several FRET biosensors (yellow cameleon...fluorescent protein are relatively close together (figure 1). Where should the fluorescent protein be inserted...fluorescent protein database." Nature methods (2019): 1. PubMed PMID: 30886412. Mastop, Marieke, et al. "A...any DNA sequence in the way that they like, the design and engineering of fusion proteins is relatively...fusion proteins and highlight some aspects of their design.  Protein size and shape matters The green fluorescent...necessary for post-translational modifications. Despite these potential issues, GFP has been used successfully...consist of random amino acid sequences or (ii) designed to form an inert, unstructured peptide and therefore...

  20. Introducing the Addgene Antibodies 101 eBook!

    Type
    Blog Post
    Published
    Oct. 29, 2024, 1:15 p.m.
    ...may find “Chapter 1: What is an Antibody” quite useful. If you’re looking to design your first flow cytometry...eBooks, the Antibodies 101 eBook is designed to be a helpful desktop reference. Each chapter brings together...
Showing: 281 - 300 of 587 results