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  1. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

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    Blog Post
    ...Scientists can also use electroporation, the application of an electrical charge to cells, to increase... properties. The phagemids are packaged into replication-incompetent phage particles with assistance from... found on an episome, a piece of DNA that can replicate on its own or be integrated within a bacterial...
  2. Antibodies 101: Monoclonal Antibodies

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    Blog Post
    ...cells and antibody-producing B cells that cannot replicate, meaning that only hybridomas will survive this... Antibodies: Distinguishing Characteristics, Applications, and Information Resources. ILAR Journal 46:... the Addgene blog Learn more about research applications for antibodies in Intro to Antibodies Read about...
  3. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

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    Blog Post
    ... specific DNA base changes and consist of a catalytically impaired Cas protein (dCas or Cas nickase) fused... SpCas9. In order to expand the gene-editing applications and targeting scope of ABEs David Liu’s lab ...through many rounds of mutations, selection, and replications per day.  The PACE and PANCE systems use M13...
  4. Building and Validating Recombinant Antibodies: A Case Study

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    Blog Post
    ...cDNA, which  was used as a template for PCR amplification with a degenerate primer set to amplify IgG ...  While screening by IF-ICC worked well for identification of the correct clones, it was determined that...through functional assays. Following functional verification, the R-mAb expression plasmids were DNA sequenced...
  5. A Novice's Perspective on Neuroscience - SfN2017

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    Blog Post
    ...the things that popped out to me as a novice and indicate why other neuroscience newbies might want to get... reinforce our goals of making more AAVs and educational content to help researchers use these great tools...start making the AAV, or help us produce new educational content on the blog. Overall, there are many ...
  6. Kiran Musunuru on the Newest TALEN Genome-Editing System

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    Blog Post
    ...Musunuru: The kit includes 832 plasmids containing prefabricated pieces that allow you to build a DNA-binding...diseases. We are knocking out genes that have been implicated in disease or, in cases where particular variants...less than a month's time. Addgene: What are the implications of this kit in terms of understanding human ...
  7. Addgene: The First Twenty Years

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    Blog Post
    ...as well as developing and sharing supportive educational resources. By 2017, we had distributed 1,000 ...We also created a growing library of antibody educational resources, including our new Antibodies 101 eBook...community by depositing, ordering, or using our educational resources. And we look forward to twenty more...
  8. CRISPR 101: Cas9 vs. The Other Cas(s)

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    Blog Post
    ...a defined site to form a clean DNA break, Cas3 locates its target sequence and initiates single stranded... out large regions of DNA and has anti-viral application potential as a result. Fast facts – PAM requirement... has been proposed to be useful for the same applications (RNA knockdown, RNA editing, etc.). The main...
  9. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

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    Blog Post
    ...sequences via oligo synthesis using the format indicated below and pool-clone them into the pAWp28 storage...realization of a variety of perturbations and applications in functional genomics, cell reprogramming, ...CRISPR/Optogentics Tools Resources on Addgene.org Catch up on Your CRISPR Background with Our CRISPR Guide...
  10. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...it is important to carefully select the target locations of knock-in studies to avoid off-target effects...it comes to targeting precision and practical application. If the CRISPR/Cas9 system is not effective for...lncRNAs with high specificity, particularly in applications requiring smaller delivery systems (Zibitt et...
  11. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...ampicillin resistance cassette and an origin of replication, which are not schematized. Sizes of fragments...five months. When they tested those clones (in duplicate), 93% had the correct banding pattern in the restriction...correct. Because they created the assemblies in duplicate, they were able to confirm that at least one plasmid...
  12. Plasmids 101: FLEx Vectors

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    Blog Post
    ...sites directs the Cre recombinase to invert, translocate, or excise a DNA fragment. The availability of... the function of various point mutations and truncations in the absence of the endogenous gene. Spatio-temporal..., allowing for clearer characterization of the location, morphology, and circuit mapping of a targeted...
  13. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
    ...-humanized (Plasmid #44246) Expression of a catalytically inactive (dead), human codon-optimized Cas9 ...be used to construct complex lentiviruses for modification of mammalian cells with a single viral infection...Gammaretroviral Vectors: Biology, Technology and Application. Viruses. 2011. PubMed PMID: 21994751. PubMed...
  14. Hot Plasmids and Viral Preps - January 2021

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    Blog Post
    ...including excision or insertion, inversion, translocation or cassette exchange. By utilizing recombinases...BioRender.com In their recent ACS Synthetic Biology publication, Mary Dunlop’s lab generated a photosensitive...regions with Vivid (Vvd) photodimers. Upon the application of blue light, Vvd dimerizes bringing the C-terminal...
  15. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ...passionate about HIV cure research and science communication.     References 1. Gaspar, H. B., S. Howe, and...affinity-purified receptor-targeted viral vectors." Nature communications 6 (2015): ncomms7246. PubMed PMID: 25665714...our AAV Production Protocol Check out our AAV Purification Protocol ...
  16. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    ... about making sure the audience is interested, educated and talking as a result of the content. Here are...contribute. The moderator should identify the funny, provocative or interesting stories that will make for a fun...sure all your panelists have the correct date, location and parking information and remind them about ...
  17. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ...This reduces the chance of integration at random locations that could cause deleterious effects. It is important...multi-plasmid system, where you have one AAV vector dedicated to expressing Cas9, while the other houses the...integration into CRISPR-induced DNA breaks. Nature Communications, 10(1). https://doi.org/10.1038/s41467-019-...
  18. Antibodies 101: Conventional vs Spectral Flow Cytometry

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    Blog Post
    ... tumors, and moreso than other methods, the identification and isolation of rare cell types. In recent...complexity index ranges from 0-1, with a value of “0” indicating those markers are completely distinct from one...one another, and a value of “1” indicating those markers are identical and completely indistinguishable...
  19. Of Myc and Men

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    Blog Post
    ...used in way more applications.   Fig. 1 Making a Protein Tag (Abridged) (1) Locate a promising monoclonal... by its antibody without additional chemical modification steps and when the epitope was within the tagged...
  20. AAV Vector Quality Control: Going the Extra Mile with NGS

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    Blog Post
    ...These impurities cannot be removed during the purification step because they are inside the virus itself...very low concentration. So how do we make sure to catch all of these potential mishaps before distributing...Guerin et al., 2020).  In addition to genome identification, we developed custom open-access Python scripts...
Showing: 701 - 720 of 1022 results