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Showing: 61 - 80 of 140 results
  1. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...cells; however, Mycoplasma species do not possess cell walls. As a result, Mycoplasma are intrinsically...fantastic example of intrinsic resistance would be Mycoplasma species’ resistance to beta-lactam antibiotics...
  2. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ... ideal number. First you generate four unique kanamycin-resistant plasmids, each containing a different...construct, you clone all but one of your gRNAs into spectinomycin-resistant plasmids pX330S-2 to pX330S-(last ...monocot and dicot plants, with a choice of either hygromycin or Basta selection. Yang Lab Single Transcript...
  3. Colony Formation Titering Assay

    Type
    Protocol
    ...obtaining accurate titers. Check the cells for mycoplasma regularly Do not over or under-grow your cells... after 20-30 passages. Do not add penicillin/streptomycin to the media. Titer will vary between cell lines... . The following day, cells were treated with puromycin. Colonies were allowed to grow in the presence...
  4. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ... The strategy relies on several observations: Rapamycin is a membrane-permeable pharmacological agent ...induction heterodimerization of proteins that contain rapamycin-binding domains, such as FKBP and FRB. The cell...membrane protein with a FRB domain (MitoTRAP). Rapamycin binds your recombinant protein (via FKBP) and ...
  5. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    ...stably integrate a reporter gene that contains a puromycin resistance gene followed by the target site (the...genome edit:  A donor plasmid that contains a puromycin resistance gene linked to an in-frame H2B-GFP....
  6. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...primarily pertains to ORIs in budding yeast, Saccharomyces cerevisiae; however, we’ve also noted some features...required for the replication of fission yeast, Schizosaccharomyces pombe, vectors at the end.) In a previous...
  7. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...deliver mammalian antibiotic resistance (e.g., puromycin, blasticidin), which enables selection of a stable...preparation Appropriate antibiotic for selection (e.g. puromycin, blasticidin) Reagent Preparation DMEM Complete...obtaining accurate results. Check the cells for mycoplasma (Link opens in a new window) regularly Do not... after 20-30 passages. Do not add penicillin/streptomycin to the media. Infection efficiency will vary...
  8. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...- and locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence...cassette into the FBL locus. Upon sequencing of puromycin resistant clones, they found that 80% and 50% ...
  9. Pouring LB Agar Plates

    Type
    Protocol
    ... µg/mL Coumermycin 25 mg/mL (dissolve in DMSO) 25 µg/mL Gentamycin 10 mg/mL 10 µg/mL Kanamycin 50 mg/mL.../mL 50 µg/mL Spectinomycin 50 mg/mL 50 µg/mL Tetracycline 10 mg/mL 10 µg/mL Note: Unless otherwise indicated...
  10. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...toxic or insoluble proteins Origami2 (DE3)** Streptomycin and Tetracycline Contains highly active thioredoxin... General protein expression  m15 pREP4*, ** Kanamycin (pREP4) Cis-repression of the E. coli T5 promoter...
  11. Five Popular Model Organisms

    Type
    Blog Post
    ... of the fly (Pfeiffer et al., 2010). Yeast (Saccharomyces cerevisiae) Yeast, one of the simplest eukaryotic....5 (2011): 190-195. Liu, Wei, et al. "From Saccharomyces cerevisiae to human: The important gene co‑expression...
  12. Fluorescence Titering Assay

    Type
    Protocol
    ...obtaining accurate titers. Check the cells for mycoplasma (Link opens in a new window) regularly Do not... after 20–30 passages. Do not add penicillin/streptomycin to the media. Titer will vary between cell lines...
Showing: 61 - 80 of 140 results