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We narrowed to 730 results for: NSI

Showing: 181 - 200 of 730 results

  1. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...immune system directly). It may seem odder when you consider that out of the 6 x 106 virus-like particles that...DNA called a “provirus.” This DNA is synthesized inside of what is called the “target cell” (or host cell...reverse transcriptase are packaged (Wu et al., 1997) inside of the capsid . This is useful for viral reproduction... particle, some of the RT protein gets packaged inside of the vector particle, along with its RT-deficient... natural virus, this new chromosomal locus is considered “proviral DNA” (Rowe et al., 1971) and the full...

  2. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    Published
    Feb. 6, 2020, 3 p.m.
    ...of current new and exciting literature. As I transitioned out of academia and away from the bench I certainly... figure organization with a different person responsible for each figure. Others still are more casual...everyone to offer thoughts you may not have even considered. These questions should be tailored to the group... not actively participating, it may be time to consider ending the meeting. This is a common failing in...participants. If you find yourself in this situation, considering stopping (temporarily or permanently) the recurring...

  3. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...immunoprecipitation (locus-specific ChIP). These methods consist of insertional chromatin immunoprecipitation (iChIP...Advisory Board.  Addgene: Your lab has worked extensively with enChIP systems. Can you describe this technology...technologies. The locus-specific ChIP technologies consisting of iChIP and enChIP are recent examples. We also...community is a kind of duty for researchers when considering the peer-review system of journals and grant ...Identification of Proteins Associated with an IFNgamma-Responsive Promoter by a Retroviral Expression System for...

  4. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    Published
    March 20, 2014, 3:04 p.m.
    ...and it will teach them to respect their own responsibilities. Another tip: don’t miss your grandfather’... the evening, or in a few years) when life responsibilities are less You can work at home sometimes, but... the burden alone You, alone, are not solely responsible for care and attention to your home and family...doctor without your help. Your partner is equally responsible and capable. The only exception is that men cannot...tasks – try to resist the impulse to control by insisting things always be done your way. Be patient with...

  5. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    Published
    Aug. 14, 2018, 12:42 p.m.
    ...AAV delivered genes. AAV serotypes have been extensively studied in order to define those that target ...weakly expressed. As these epitopes tags have no intrinsic visual signal, they cannot be detected directly...neurons. As compared to epitopes tags, FPs are intrinsically fluorescent and can therefore be used in live...darkened chromophores. These probes are both intrinsically fluorescent and can be detected with highly ...tags like GFP. Specifically for STORM, the high intensity signal provided by smFPs can result in better ...

  6. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    Published
    Jan. 17, 2019, 1:38 p.m.
    ...emerged from intensive engagement with a wide range of people at the University of Wisconsin in 2011/12 ...reproducibility is limited in two ways – only data inside the ELN is reproducible, and only the highly restricted...principles emerging from the engagement with the Wisconsin research community were to: (1) prevent vendor... choosing an ELN for your research, be sure to consider how its features do and don’t support reproducibility...

  7. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ... package stays in transit for a certain number of days and tracking average transit time for each courier...having some materials available to order, yet not consistently accessible, wasn’t an option. So we identified...For example, when we began offering temperature-sensitive materials, we noticed a large percentage of these...distributor allowed us to ensure that temperature-sensitive materials like viral preps and libraries were ...

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

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ...Specificity concerns? If off-target effects are a worry, consider a dual-sgRNA system. This system uses a “nicking...little homology to other regions of the genome and consider using a Cas9 variant with higher specificity (...requirements, giving it more flexibility. A consideration with this tool is that the desired edit will...activators The simplest of the dCas9 activators consists of dCas9 fused to a single transcriptional activation...Mediator complex, generally the strongest activator, consists of a dCas9-VP64 fusion and a modified gRNA that...

  9. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    Published
    Nov. 15, 2017, 3 p.m.
    ...viral vector solution. Since this technique is intensive and requires an electron microscope, it is not...image shows that the vast majority of the vectors consist of full particles (white arrowhead) relative to...from different sources in the same experiment, consider retitering both AAV in your lab. The absolute ...does not give an indication of infectious titer, consider titrating each lot of AAV in vivo to determine...Additional Resources on the Addgene Blog Important Considerations When Using AAVs Addgene's AAV Quality Control...

  10. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    Published
    Nov. 10, 2016, 3:30 p.m.
    ...resistant to antibacterial drugs. Resistance can be intrinsic, meaning a basic characteristic that naturally...resistant somehow overtime. One fantastic example of intrinsic resistance would be Mycoplasma species’ resistance...possess cell walls. As a result, Mycoplasma are intrinsically resistant to this class of antibiotic. Acquired...be unharmed by the compound it produces (think intrinsic resistance), but surrounding microbes would be...prescribed to them This may seem counter-intuitive considering the other three scenarios, but a short treatment...

  11. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...sure your cells have first reached an appropriate density. Cells in the exponential growth phase are alive... prevent the culture from reaching the desired density within a reasonable time-frame. For these cases... growth media, or if your plasmid is low-copy, consider reducing the antibiotic concentration. Growth ...accomplished by growing your culture to the desired density at 37°C and reducing the temperature or moving ...TB or 2XYT is optimal because of the high cell-density they support; however, minimal media supplemented...

  12. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...gene of interest) and prevent incorporation of extensive sequences used for cloning into the final translated...Biosafety of biotechnologically important microalgae: intrinsic suicide switch implementation in cyanobacterium... integrity, expression level, and aggregation propensity in four expression systems: E. coli, wheat germ...frequencies across timepoints or populations. This kit consists of a 48 plasmid adapter library, with each plasmid...Morgan’s blog post on the PUF Assembly Kit for some insights on its potential from its creators. We hope you...

  13. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...concentration of Taq polymerase, 2) increased PCR extension time, 3) increased concentration of MgCl2, 4) ...vector with Gibson assembly. While this is a comprehensive mutagenesis method, a drawback to using Gibson...scanning libraries. PFunkel: Of prior existing comprehensive mutagenesis methods, PFunkel offers the best...PCR-based approach for generating a customizable comprehensive mutagenesis library that’s ready to be tested...Elad, and Marc Ostermeier. “PFunkel: Efficient, Expansive, User-Defined Mutagenesis.” Ed. D. Dafydd Jones...

  14. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ...antibodies, increasing the fluorescence signal intensity. However, you need to be careful when selecting...interest is not located on the cell membrane but inside the cell. Due to their chemical nature, antibodies...nucleus intact. If markers of interest are located inside the nucleus, a different fix/perm kit is required...that can enter cells freely are typically used. Inside the cell, the ester group is cleaved off by enzymes... yielding a stepwise decrease in fluorescence intensity in a (histogram) flow plot (Figure 4b). Through...

  15. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    Published
    Sept. 29, 2020, 1:15 p.m.
    ...metabolic engineering.  The E. coli genome has an extensive research history and characterization. Many proteins...cardiovascular disease, specifically stroke and hypertension (Iannaccone et al., 2009). Rodents have also...amphibians in science with early research providing insight into some of the most significant and fundamental... plant model organisms in the world. With its extensive scientific history, highly characterized genome...Genome sequence of the Brown Norway rat yields insights into mammalian evolution. Nature 428, 493–521 ...

  16. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...included inducible Cre, such as the tamoxifen-responsive CreERT, the coupling of Cre-lox with FLP-FRT ...limited due to low resolution or were very time-intensive, necessitating the development of a new technique...This system differs from Brainbow-1.0 in that transient Cre recombination is required to prevent constant...tissue was challenging due to low fluorescence intensity, caused partially by fluorophore photoinstability... the Brainbow technique will provide important insights into the complicated physical organization of ...

  17. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...prokaryotic minions that carried out the labor-intensive process of replicating your new plasmid: the Escherichia...certain restriction enzymes that are methylation sensitive. dnaJ Mutation in a chaperonin gene Increases...strains are all based on E. coli K-12 and are considered BSL-1. Table 2: Lab strains of E. coli Strain...Dam or Dcm methylated, allows for methylation sensitive restriction enzymes to cut the plasmid after preparation...tables are by no means exhaustive! For a more comprehensive list and additional information, visit OpenWetWare's...

  18. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Sept. 24, 2020, 1:15 p.m.
    ...gRNAs! Comic by Maya Kostman.   Important considerations before you start an experiment with CRISPR ...expression. Location and sequence are important considerations for designing your gRNAs. For indels, it's ...off targets. For CRISPRa and CRISPRi, these considerations are of roughly equal importance (target should..., 2020). is not limited to single nucleotide transitions but still requires a nearby PAM, although these...The off-target activity of gRNAs is important to consider. While the basic landscape of mismatches that ...

  19. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    Published
    Nov. 2, 2021, 1:15 p.m.
    ...FLEx Vectors Important Considerations When Using AAVs Three Key Considerations For Precise Neuronal Targeting...-target expression.  Some classes of neurons transiently express TH during development or have low levels...titer to prevent off-target expression of the sensitive FLEx-split TVA receptor used for tracing neural...recombinase-dependent AAV vectors and mitigation with cross-over insensitive ATG-out vectors. Proc Natl Acad Sci USA 116:27001...

  20. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...the Moloney Murine leukemia virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing...vectors Although retroviruses are still in use, considerable efforts have been devoted to develop efficient...generation. The 1st generation of lentiviral vectors consisted of three plasmids encoding 1) the lentiviral vector... from one batch to another. For experimental consistency it’s a good idea to purify and titer your virus...sucrose to the supernatant to create a gradient of density that favours virus precipitation. My tip for this...
Showing: 181 - 200 of 730 results