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We narrowed to 736 results for: RAN-1

Showing: 361 - 380 of 736 results

  1. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    ...our lives. Plasmid transformation into E. coli is a fairly inefficient process– just 1 out of 10,000 cells...down especially fast and plates should be used witin 1 month for optimal efficiency. Beware of satellite ...aminoglycoside Binds 30S ribosomal subunit; causes mis-translation Bactericidal 50-100 ug/mL Spectinomycin aminoglycoside...Blocks 50S ribosomal subunit; inhibits aminoacyl translocation Bacteriostatic 50-100 ug/mL in EtOH Polymyxin...Polymyxin B polypeptide Alters outer membrane permeability Bactericidal 10-100 ug/mL Tetracycline tetracyclin...Binds 50S ribosomal subunit; inhibits peptidyl translocation Bacteriostatic 5-25 ug/mL in EtOH *In prokaryotes...

  2. CRISPR Challenges: Standardization and Homology Directed Repair

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    Blog Post
    ...B. Gurumurthy, and Masato Ohtsuka. Nat Protoc. 13(1) (2018):195-215. PubMed PMID: 29266098 Creating knock-in..., with protocols using dsDNA templates achieving ~1-10% efficiency. Easi-CRISPR uses a long ssDNA template...sub-pathways like synthesis-dependent strand annealing (SDSA) and single-strand DNA incorporation (ssDI). "Easi-CRISPR...These complementary goals are aimed at improving translation of CRISPR technology, but they would also bring...a DNA repair newbie - diagrams of resection and strand invasion are like a foreign language to me. But...supports over 250 genomes, and they just released a brand new CRISPOR manual to kick off 2018! "Mechanisms...

  3. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

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    Blog Post
    ...more tools for plasmid mapping and design (Figure 1). Benchling: While you might think of Benchling as...Davis from the University of Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid...sequences to sequences in a database ExPASy translation tool: Translates a DNA sequence using all six open reading...Annotates features, BLASTs, aligns, digests, and translates a sequence.   NEBCutter2: For finding restriction...information, such as expression patterns, post-translational modifications and polymorphisms Codon optimization...

  4. Hot Plasmids and Viral Preps - July 2021

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    Blog Post
    ...organoid model (Sun, et al. 2021).  Figure 1: Example schematic from the Organoid EasyTag system...without altering that of the internal control (Fig 1). Because this method relies on the fluorescence ratio...B) Strategy for leak-free transcriptional control. C) Iterative transduction workflow overview. Image ... by integrating a SL-insensitive fluorophore (LSSmOrange) as an internal control and a SL-sensitive fluorophore...fluorescent biosensor has a high sensitivity (50 nM-500nM range) and a high specificity for SLs in vitro, and they...

  5. Year of the Camelid: Antibody Style

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    Blog Post
    ... similar antibodies in llamas and alpacas (Figure 1). At the time, researchers were already on a quest...nanobodies (Arbabi-Ghahroudi, 2017)!      Figure 1: Comparison of a heavy-chain only antibody and single-domain... tools for diverse applications. For example, RANbodies can be used as an alternative to conventional ...sdAbs were used to create GFP scaffolds for transcriptional activation.  sdAbs in the clinic sdAbs have...also been some interesting work done developing transgenic mice that express camelid variable domains (aka...Llamas Required - Synthetic Nanobodies Against Membrane Proteins More resources from addgene.org  Plasmids...

  6. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    ...cleavage. NgAgo takes a different approach, randomly removing 1-20 nucleotides from the cleavage site specified...About NgAgo User's First Impressions Here References 1. Gao, Feng, Xiao Z. Shen, Feng Jiang, Yongqiang Wu...in the cell. gDNA guides will also need to be transfected into the cells of interest. In contrast, RNA ...must be expressed from a plasmid or in vitro transcribed, and changes in gRNA secondary structure can ...removal RNA-guided CRISPR enzymes cause double-stranded breaks, with either blunt (Cas9) or staggered ...

  7. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

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    Blog Post
    ...from distantly related organisms. In 2006, less than 1% of the genes deposited at Addgene were synthetic....synthetic biology is expanding rapidly.  References 1. Kunjapur, Aditya M., Philipp Pfingstag, and Neil ... synthesis companies often work within a narrow range for GC content. Rare codons also are not good predictors...sequences is Proteobacteria - the most frequent transfer of genes is from Proteobacteria to mammalian expression...natural gene sequence is, the less likely it is transferred into another organism in the lab. Predicting ...

  8. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

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    Blog Post
    ...optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference... plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan, A., ...expressing ECN is 109 cells/ml, which is within the range of  the ~1011 commensal bacteria per gram of stool...

  9. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

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    Blog Post
    ...receptors. Biochimica et Biophysica Acta - Biomembranes, 1838(1), 15–33. https://doi.org/10.1016/j.bbamem...principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes... dopamine D2/D3 receptors. Scientific Reports, 10(1), 21842. https://doi.org/10.1038/s41598-020-78827-...for bacterial imaging . Microbiology Spectrum, 12(1), e0369023. https://doi.org/10.1128/spectrum.03690... of fluorescence: For applications that involve ranges of pH, and significant lengths of time, the stability...receptor conformation and signaling activity​​ (Sridharan et al., 2014). It's also crucial to evaluate how...the background. Dyes that can pass through cell membranes can prove useful for live-cell imaging. Experimental...

  10. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

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    Blog Post
    ...this system.  Fig. 1: Cytidine deamination takes place on the free strand of DNA and converts a C...guide RNA (sgRNA), provided it meets two conditions:  1) The sequence is unique compared to the rest of the...temperature-controlled genome editing. Nat. Commun. 2017 81 8, 1–9 (2017).Walton, R. T., Christie, K. A., Whittaker...changing its double-strand cut to a single-stranded nick. In order to produce a double stranded break, two sgRNAs... target DNA. The reverse transcriptase polymerizes DNA onto the target strand based on the sequence of...targeting dCas9 to transcriptional start sites was sufficient to repress transcription by blocking its initiation...approach is that two sgRNAs targeting opposite strands must be available in moderate proximity to each...

  11. The Advances Behind The World's First Personalized CRISPR Treatment

    Type
    Blog Post
    ...  References and Resources References Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N.... Cell, 164(1), 18–28. https://doi.org/10.1016/j.cell.2015.12.041  Musunuru, K., Grandinette, S. A., Wang...may contribute to more in the future. Figure 1: A timeline of CRISPR base editing advances that led...the safety of their intervention, the team led by Kiran Musunuru and Rebecca Ahrens-Nicklas administered...disorder survive long enough to receive a liver transplant — creating intense time pressure for the research...al., 2017). These editors did not create double-stranded breaks in the DNA backbone, making them significantly... repaired with an A→G edit on the complementary strand. But the invention of base editing doesn’t necessarily...

  12. Plasmids 101: Plasmid Incompatibility

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    Blog Post
    ... RNAs inhibit plasmid replication.   Figure 1: The replication problem. Iterons are repeated ...actually does, and neither plasmid is maintained (Figure 1).  The partitioning problem Plasmid incompatibility...Agrobacterium tumefaciens. Genes & Genetic Systems 77:1–9 . https://doi.org/10.1266/ggs.77.1 Pubmed PMID: ... and inhibit its own transcription. As copy number increases, RepA transcription is repressed and replication...indirectly inhibit replication through inhibiting translation of replication machinery proteins, or directly... division. They don’t simply rely on chance (or random diffusion) to make this happen.  There are several...CBPs and NTPase to go around, the plasmids are randomly positioned, leading to plasmid loss (Diaz et al...

  13. Hot Plasmids: Fall 2024

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    Blog Post
    ...and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody or want...are also available from Addgene!   Figure 1: Experimental validation for Anti-V5 [IPI-SV5-Pk1]...biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https://doi.org/10.1038/s41467-024-52871-9... the P and V proteins of Simian virus 5 (SV5) (Randall et al., 1987). The epitope (GKPIPNPLLGLDST) has...10.57733/addgene.l0ulu4. Hanke, T., Szawlowski, P., & Randall, R. E. (1992). Construction of solid matrix-antibody-antigen....org/10.1099/0022-1317-73-3-653. PMID 1372038. Randall, R. E., Young, D. F., Goswami, K. K., & Russell...functional properties, and is amenable to a wide range of biophysical assays (Kroupova et al., 2024). They...

  14. Tips for Getting a Faculty Position

    Type
    Blog Post
    ...sentences) What will be the focus of your first grant? (1-2 sentences) Simply stated, what are your...Full "Applying for a Faculty Position" eBook Here 1. Have a competitive CV You don't necessarily need ...that people would agree is important to understand. (1-2 sentences). For example: Distinguishing correctly...emphatically deny that I have a magic formula to guarantee a reader the faculty job of his/her dreams. There...currency of scientific success is publications and grants. 2. Write succinct and easy to understand documents...years. You will discuss the focus of your first grant, how you will overcome certain obstacles, alternative...It's not about you and how you will finally be granted the faculty position you have craved since grad...

  16. Hot Plasmids - October 2022

    Type
    Blog Post
    ...protein sequence of the predecessor GEVIs (called ASAP 1 and 2), they found an indicator that is optimized ...several other constructs in plasmid form. Fig. 1: Summary of the approach and outcomes of the screen...voltage indicator: JEDI-2P  by: Brian O'Neill  The François St-Pierre lab and collaborators have successfully...photoactivation of HcKCR1 in mouse neurons. D) Membrane voltage traces of action potentials in a HcKCR1...

  17. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...screening for the best gRNA Figure 1: Validation of double strand break (DSB) mediated homology dependent...as control (Cetn1/sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence indicates a more...and greatest in scientific content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target effects...repair (HR, homologous recombination; SSA, single strand annealing) took place and reconstituted the EGFP...site. (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences...efficacy of your gRNAs is then a simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP...fluorescence, the better that gRNA is at causing a double stranded break at the target site, allowing the two halves...

  18. Important Considerations in Optogenetics Behavioral Experiments

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    Blog Post
    ...derekpsimonphd.    References 1. Stuber, Garret D., et al. "Excitatory transmission from the amygdala to nucleus...animal will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models... can be explained primarily in two different ways 1) your hypothesis is wrong or 2) there is some problem...examples I’m personally interested in include: intracranial self-stimulation (ICSS) and place preference...(viral expression, efficiency of laser light transmission to the target neurons, appropriate stimulation...optogenetics is performed in this species. However, transgenic rat technology is improving and may one day reach... reach the sophistication of mouse transgenics. Everything that has been described is intended for the...

  19. Allen Institute for Brain Science AAV Enhancer Collection

    Type
    Collection
    ...Tools Atlas (Link opens in a new window) . Figure 1: Selection, prep, and testing of AAV enhancers in ...Tasic, B. (2025). A suite of enhancer AAVs and transgenic mouse lines for genetic access to cortical cell...

  20. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

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    Blog Post
    ...References 1. Bressan RB, Dewari PS, Kalantzaki M, Gangoso E, Matjusaitis M, Garcia-Diaz C, Blin C, Grant V, ...the tag sequence flanked by homology arms (Figure 1). In the first step of optimization, we started off...form the ribonucleoprotein complex (csRNP, Figure 1). We use in-house purified Cas9 protein, which, in...standard immunocytochemistry (ICC) assays.  Figure 1: Three ingredients of epitope-tagging experiments....binding to the template strand (strand used by RNA polymerase II for transcription) are more effective for...complex. This complete transfection mix, i.e. csRNP plus ssODN, is then transfected into stem cells. Cells... were transfected in 96-well format using an Amaxa Shuttle device; 5 days after the transfection, the ...
Showing: 361 - 380 of 736 results