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We narrowed to 961 results for: Cre

Showing: 581 - 600 of 961 results

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

    Type
    Blog Post
    ...primary antibody and endogenous IgGs in the tissue. Created with biorender.com. Strategies for avoiding...Some labs have found that you can significantly decrease background staining by mixing your primary and...you add your primary and secondary antibodies. Created with biorender.com. Proper tissue processing...

  2. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    ...Recombinant antibodies are plasmid-based and are created synthetically or derived from hybridoma antibody...antibodies against the new Fc region. In addition to increasing multiplexing capabilities this process is frequently...Although isotype conversion is routinely used to increase the utility of recombinant antibody tools, the...

  3. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ...single-strand DNA incorporation (ssDI). "Easi-CRISPR for creating knock-in and conditional knockout mouse models...Protoc. 13(1) (2018):195-215. PubMed PMID: 29266098 Creating knock-in mouse models with CRISPR has been challenging...efficiency. Easi-CRISPR uses a long ssDNA template to increase efficiency of insertion and gene replacement. ...

  4. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    ...we found that adding a TEV linker dramatically increased accessibility of the AviTag and HisTag, which ...fusion protein in bacteria. Our initial attempt to create a plasmid based on the biotin ligase BirA backbone... inserting BirA into a pMal plasmid (NEB) thus creating the pMal-T-Avi-His/BirA plasmid now available ...

  5. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ...individual cells. Each cell is labeled with 3-4 tags to increase the diversity of the CellTag library. The fixed...detected over a 10-week period. Their expression decreased over time but they were not silenced.    Find ...retroviral overexpression cocktail yielded a three-fold increase of fully reprogrammed cells based on single-cell...

  6. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    ...University of Alberta and University of Oregon created mTFP1 by transforming E. coli with a gene library...tetramer-breaking mutations. The library was then screened through successive rounds of mutagenesis for the...filter. This combination results in a 2.6-fold increase in signal relative to mCerulean imaged with a ...

  7. RUBY-Red Siliques

    Type
    Blog Post
    ...to select against CRISPR positive plants, but screening these can be challenging and requires specialized...Willoughby   Using RUBY as a selection marker The creators of RUBY highlight its use in crop plants under...as positive selection of transformed plants to decrease the routine usage of antibiotics which may contribute...

  8. Plasmids 101: Origin of Replication

    Type
    Blog Post
    ...R6K, and 15A. Not all origins of replication are created equal. Some will produce many plasmid copies and...because they will compete for the same machinery, creating an unstable and unpredictable environment. As ...plasmids if they contain large inserts or genes that create a toxic product. The E. coli strain Most E....

  9. Summer SciComm: Science Storytelling

    Type
    Blog Post
    ... - not just for fiction! Image credit: Kenneth Rowe, under the creative commons license.    The introduction...is to find one or two roadblocks that sparked creativity, changed the outcome of the project, or illustrates...

  10. Addgene Begins Distribution of Recombinant Antibodies

    Type
    Blog Post
    ...collection of antibodies available at launch were created by James Trimmer, PhD, of the University of California...commitment to supporting your research, Addgene is creating an array of educational materials to accompany...more useful to everyone over time. Through our incredible community, including your support, we have enabled...

  12. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    ...cassette allowing a researcher to perform blue-white screening after transformation - self joining of the vector...break down the steps needed for TOPO cloning: 1. Create Your PCR Product: Design standard primers (no need...unpaired 3’ ends in your PCR product. If you need to decrease error rate, please use one of these methods to...

  13. Don’t Wait To Launch Your Health Science Startup

    Type
    Blog Post
    ...would rather fund excellent people with mediocre ideas than mediocre people with excellent ideas. I am lucky... your potential as an entrepreneur, not your credentials. If you have an idea, don’t wait.  Go for it....

  14. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ...microvesicles (approx. 40 - 150 nm diameter) that are secreted from the cellular membrane into the culture medium...AAVs (Schiller et al., 2018). Exosomal vesicles secreted from producer cells also contain cellular macromolecules...open new ways of improving gene delivery. They increase the efficiency of transduction, broaden the use...

  15. Virtual Science Conference Coming Up? Three Addgenies Share Their Experience

    Type
    Blog Post
    ...to keep the audience engaged through a computer screen. For the Reproducibility for Everyone Workshop,...author of the poster,” he says.Viewing a poster on a screen is challenging, requiring you to zoom in to different...canceled may be disappointing, but there are many creative ways to adapt and take advantage of the new conference...

  16. Four Factors that Differentiate the Stem Cell Field

    Type
    Blog Post
    ... the stem cell community – a community largely created due to Yamanaka’s seminal work. It didn't take... during his keynote address, appeared almost incredulous that the field has exploded so quickly, citing...explained that it took 50 people nearly 2 years to create the first stem cell line that could be used in ...

  17. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    ..., described below, are typically employed when creating stable cell lines and are not used for general...Stable transfection A stable transfection is used to create a population of cells that have fully and successfully...culture media until 24-48 h post transfection when creating stable cell lines. Looking for Mammalian Expression...

  18. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ... vector and insert are (separately) digested to create 10–12 base “chewed back” overhangs, they can be...bacteria, which will efficiently repair the plasmid. Created with BioRender.com.   SLIC also opened up a...transformed into bacteria, which repair the plasmid. Created with BioRender.com. How does SLIC compare to...

  19. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...sequence (pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP ...virus, making it ideal for CRISPR pooled library screening experiments. Find lentiCRISPR v2.(If you’ve peeked...site, a pair of annealed oligos can be cloned to create a chimeric guide RNA. Find pX330-U6-Chimeric_BB-CBh-hSpCas9...

  20. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    ...under cold temperatures, and frigid conditions decrease the potential for microbial contamination. Additionally...Acclimatisation To resolve this impasse, our team decided to create a hybrid system where we could incorporate cold-tolerance...The event also featured a seminar on “Tools to increase scientific reproducibility” by Benoit Giquel, ...
Showing: 581 - 600 of 961 results