We narrowed to 144 results for: MYC;
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TypeBlog Post...but was found to have a point mutation in the puromycin cassette that makes it less efficient in some ...with customizable sgRNA from the U6 promoter and puromycin resistance from the EF-1a promoter. It’s a 3rd...PX459) V2.0. It contains a point mutation in the puromycin cassette that makes it less efficient in some ...
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Protocol - pLKO.1 – TRC Cloning Vector
TypeProtocol...kinase promoter drives expression of puromycin. Puro R Puromycin resistance gene for selection of pLKO...sulfate, and puromycin are located in the “Appendix”. F.2. Determining the Optimal Puromycin Concentration...differently to puromycin selection. Addgene strongly recommends that you determine the optimal puromycin concentration...to fresh puromycin-containing media every other day. f. The minimum concentration of puromycin that results...with finer increments of puromycin to determine a more precise optimal puromycin concentration.) F.3. Protocol...expression of the puromycin resistance gene. f. To select for infected cells, add puromycin to the media at...positive control for the puromycin selection. Days 4+: g. Change to fresh puromycin-containing media as needed... -
Visualizing Protein Turnover In Situ
TypeBlog Post...Nathans, Daniel. "Puromycin inhibition of protein synthesis: incorporation of puromycin into peptide chains...Yarmolinsky, Michael B., and L. Gabriel. "Inhibition by puromycin of amino acid incorporation into protein." Proceedings... -
Tips for arabidopsis transformation
TypeBlog Post... plant selection markers (basta, hygromycin, kananamycin, tunicamycin) (Nakagawa et al., 2007). Alternatively... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog 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... -
CRISPR 101: Multiplex Expression of gRNAs
TypeBlog 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... -
CRISPR/Cas9 FAQs Answered!
TypeBlog Post...marker such as puromycin, you can (also) select for the positive colonies by puromycin selection. You ...transfected cells (for example, plasmid with a puromycin resistance cassette, such as PX459). After you... -
Even more elegant: Single injection CRISPR/Cas9 in C. elegans
TypeBlog Post...uses a selection cassette containing genes for hygromycin resistance and the Roller movement phenotype....young adult worms. Once the worms have laid eggs, hygromycin is added to the plates to select for candidate... -
22 Hot Plasmid Technologies from 2014
TypeBlog 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 ... -
Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2
TypeBlog 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.... -
Plasmids 101: Yeast Vectors
TypeBlog 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... -
PITChing MMEJ as an Alternative Route for Gene Editing
TypeBlog 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% ... -
Plasmids 101: E. coli Strains for Protein Expression
TypeBlog 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... -
Protocol - How to Inoculate a Bacterial Culture
TypeProtocol...Chloramphenicol 25 µg/mL Coumermycin 25 µg/mL Gentamycin 10 µg/mL Kanamycin 50 µg/mL Spectinomycin 50 µg/mL Tetracycline... -
Five Popular Model Organisms
TypeBlog 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... -
Colony Formation Titering Assay
TypeProtocol...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, as it can affect viability and... . The following day, cells were treated with puromycin. Colonies were allowed to grow in the presence... -
Virus Protocol - Generating Stable Cell Lines
TypeProtocol...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, as it can affect viability and... -
Pouring LB Agar Plates
TypeProtocol... 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 Notes: Unless otherwise indicated... -
Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins
TypeBlog Post...during the development and disintegration of mycobacterial granulomas." Immunity 28.2 (2008): 271-284. ...presentation and T cell effector function in mycobacterial granulomas." Immunity 34.5 (2011): 807-819. ... -
Fluorescence Titering Assay
TypeProtocol...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, as it can affect viability and... -
New Podcast Segment: Hot Plasmids
TypeBlog Post...this kit to complement unused auxotrophies in Saccharomyces strains. Listen to Our Segment on The Yeast... -
Tag Your Favorite Yeast Genes with Ease
TypeBlog Post...and tagging at either the N- or C-terminus (3xHA, 13xMyc, GST, or GFP). Longtine et al.(3) describe a complimentary... -
What's New in CRISPR - March 2020
TypeBlog Post...that controls repression of gene expression in Saccharomyces cerevisiae. The system consists of dCas9, the... -
pSiM24: Simplifying Plant Genetic Engineering
TypeBlog Post...functional in a variety of plant species. The kanamycin antibiotic resistance gene is controlled by a ... -
What the HEK?
TypeBlog Post...cell lines are susceptible to contamination by mycoplasma and bacteria, but HEKs are also particularly ... -
Plasmids 101: A Brief History of Plasmids and an Improved eBook!
TypeBlog Post...resistant plasmid, pSC101, and a newly developed kanamycin resistant plasmid, pSC102, with EcoRI and selected... -
Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]
TypeBlog Post... various metabolites in Escherichia coli or Saccharomyces cerevisiae with few techniques that are easily... -
K. phaffii: Rising to the Occasion
TypeBlog Post...Resources References Liu, Wei, et al., From Saccharomyces cerevisiae to human: The important gene co‑expression... -
Visualizing Translation at the Single Molecule Level
TypeBlog Post...Using translation inhibitors cycloheximide and puromycin, Halstead et al. verified that translation is ... -
Four Factors that Differentiate the Stem Cell Field
TypeBlog Post...by expressing four proteins Oct4, Sox2, Klf4, and cMyc (the so called OSKM factors). As the Yamanaka lab's...