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We narrowed to 913 results for: nes

Showing: 401 - 420 of 913 results

  1. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...plasmid yield and/or DNA quality. Table 1 below outlines a few of the more common genetic changes found...encodes proteins needed for bacterial conjugation. Genes listed on F´ are wild-type unless indicated otherwise...lac Lac operon mutations Blue/white screening of clones   lacIq lac repressor overproduced, expression...transformation efficiency deoR constitutive expression of genes for deoxyribose synthesis Allows uptake of large...)]  Note: Inactivating mutations in specific genes are signified with a minus sign (-) as is typically...

  3. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...edits and to perform functional screens to identify genes involved in a phenotype of interest. This blog post...protein-coding capacity of a locus. When using S. pyogenes Cas9 (SpCas9), potential target sites are both...Veres et al., 2014). When working with single-cell clones, the authors note that “clonal heterogeneity may...obstacle to the generation of truly isogenic cell lines than nuclease-mediated off-target effects.” Further... CRISPR-Cas9 and TALEN Targeted Human Stem Cell Clones Detected by Whole-Genome Sequencing. Cell Stem ...

  4. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...al., 2009), and at high copy numbers (Williams, Carnes, and Hodgson, 2009). More research is ongoing to...1566524020666200427110452. PMID: 32338220. Crozat E, Fournes F, Cornet F, Hallet B, Rousseau P. Resolution of...Levene SD. Analysis of DNA topoisomers, knots, and catenanes by agarose gel electrophoresis. Methods Mol Biol...0092-8674(84)90060-6. PMID: 6323019. Williams JA, Carnes AE, Hodgson CP. Plasmid DNA vaccine vector design... JA, Luke J, Langtry S, Anderson S, Hodgson CP, Carnes AE. Generic plasmid DNA production platform incorporating...

  5. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...involves making a large sgRNA library to thousands of genes and targeting and sequencing millions of cells. ...cell line, with the goal of isolating single cell clones to use for multiple downstream experiments. You...certain Cas enzymes are less compatible with the bulkiness that accompanies mispairing of bases. To reduce... characterization of, isolating 2 or 3 distinct clones to perform key experiments on is a good way to ... distinct off-target sites, eliminating its effectiveness as a control. Since off-targets typically confound...

  6. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    ...Finally, as Pamela Hines of AAAS points out on the panel, irreproducibility undermines public trust in science...Last Wednesday we worked with the Harvard GSAS Science Policy Group to organize a Minisymposium on Reproducibility...) - Panel Alex Tucker (Ginkgo Bioworks) Pamela Hines (Senior Editor at Science) Edward J. Hall (Professor...journalists and scientific experts discussed by Pamela Hines Minisymposium on Reproducibility Booklet - Including...

  7. Top Requested Plasmid of 2017 - pMD2.G

    Type
    Blog Post
    ...many of which are used to deliver Cas9 and other genes to mammalian cells. For this reason, the top requested...means researchers who use VSV-G can deliver their genes of interest to a variety of cells and study their...Skimming the list, one gets a sense of the broad usefulness of pMD2.G and VSV-G. This plasmid has been used... of the secretome, ubiquitin ligase, cellular senescence, autophagy, CRISPR, and so much more (find the...

  8. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ...societal implications even before the discovery of genes themselves.  How has scientific progress in genetic...and vice versa? We draw a parallel between the timelines of the scientific advancements and the noteworthy...to stars in space ships and the rise of killer machines that exterminate entire humanity were the two ...that results in gene knockouts.   Star Trek: Into Darkness (2013) which is the remake of the space-melodrama...Hugo De Vries: from the theory of intracellular pangenesis to the rediscovery of Mendel. Comptes Rendus ...

  10. Water Bath Protocol

    Type
    Protocol
    ...digests needed in molecular biology, and thaw cell lines. There are many different types of water baths that...

  11. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ...Purpose  SpCas9 This plasmid expresses Streptococcus pyogenes (Sp) Cas9 from a Cbh promoter. Cbh is a modified...Zhang lab has tested in a variety of mammalian cell lines. Cas9, of course, is the key component of most CRISPR...gene that made it less effective in certain cell lines. This point mutation has been corrected in V2.0....gRNA of interest in a variety of mammalian cell lines. Alternative forms of this vector contain GFP-tagged...

  12. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...visualize the protein via fluorescence or chemiluminescence, precipitate the protein out of solution, ...region   The constant region of the heavy chain determines the antibody’s isotype - that is, one of five...which come from naturally-occurring immunoglobulin genes, recombinant antibodies are plasmid-based. Their...antibody allows the protein to be detected by chemiluminescence or fluorescence. Western blots are frequently...Steven GE, Lorna J. (2004) ANTIGENS, IMMUNOGENS, VACCINES, AND IMMUNIZATION. In: Immunology Guidebook. Elsevier...

  13. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...overview of the instrumentation A flow cytometer combines three systems to analyze single cells from a mixture...degree of internal complexity such as extensive membranes produces a greater SSC.  Protein expression, measured...targets, measured by fixing and permeabilizing membranes For intracellular protein targets, users must ...propidium iodide, can only enter cells with disrupted membranes as is the case with dead or dying cells. DNA binding...the cells, amine reactive dyes interact with free amines in the cytoplasm causing dead cells to fluoresce...

  14. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

    Type
    Blog Post
    ...screens using CRISPR technology, gRNAs to knockout genes – are essential screening components, the model ...Sometimes, you might have a few positive control genes on which to test the validity of your biological... a new assay or both, and you have no idea what genes are involved. Or you don’t know how well your assay...definition, the kinome is a well-annotated collection of genes that can provide a useful starting point for developing...

  15. Editor's Choice, August 2016

    Type
    Blog Post
    ...Directed Mutagenesis by PCR Google Forums Round Up: First Impressions of NgAgo CRISPR Between the Genes: How...optimize and troubleshoot the system. Site Directed Mutagenesis by PCR & Single Base Editing with CRISPR For ...technology may make some uses of Site Directed Mutagenesis by PCR obsolete! Honorable mentions Of course...

  16. IDP and your PI: A Roadmap for Career Planning and Personal Development

    Type
    Blog Post
    ...miss out on beneficial opportunities or to find oneself unsure of the next steps. Harvard PI Angela DePace...lab culture. The aim of these meetings is to give honest, constructive feedback, set tangible research and...personal development through thoughtful planning and honest feedback DePace’s yearly planning meetings revolve...person organize his or her thoughts to facilitate honest conversation. Goals and Planning Worksheet: ...

  17. Who Gives a Tweet? 9 Facts About Scientists on Twitter

    Type
    Blog Post
    ... more often than other disciplines. The majority of tweets for most disciplines contain scientific domains...#8. Figure 1: Highly Represented Scientific Disciplines on Twitter. Twitter users were identified as ...Follower networks of subgroups of scientific disciplines: social (left), life (center), and physical (...

  18. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

    Type
    Blog Post
    ...based on a protein isolated from Streptococcus pyogenes that locks itself together, to produce irreversible...another important step forward by dissecting that S. pyogenes protein into three parts. Their efforts yielded... from the “S” in Streptococcus and the “py” in pyogenes), capable of locking two peptide tags together... chain.  Addgene: What about the Streptococcus pyogenes protein domain made it ideal for this purpose?...

  19. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    ...into the following categories: Over 300 empty backbones with 100+ distinct fluorophores to choose from...constructs for biosensing. Over 300 plasmid backbones spanning a rainbow of fluorescent proteins “It...It’s a great collection with over 300 backbones alone,” said Addgene’s Lianna Swanson, who has been working...collection with such variety.” In most cases, backbones are available to attach the gene of interest to...

  20. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ... Encodes insert(s) A plasmid with one or more genes, protein coding sequences, or other elements inserted...designed for cloning a gene/insert into it. Empty backbones often contain epitope tags or fusion proteins ...Insert *Note: This page will not appear for empty backbones Please enter the name of any gene contained within...Addgene ID in the "Backbone Manufacturer" box for backbones requested through Addgene. For example, you could...Addgene qualifies as BL1 or BL2 as defined by NIH guidelines. I hereby certify that I have disclosed all known...
Showing: 401 - 420 of 913 results