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We narrowed to 730 results for: NSI

Showing: 161 - 180 of 730 results

  1. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    Published
    March 15, 2022, 1:15 p.m.
    ...secondary considerations to validation, application specificity, and experimental considerations - no other...has been validated in your specific application. Consider also that there may be variations within an application... your needs.    Multiplex experiments Finally, consider whether you will be using other antibodies in ...from a vendor that you use frequently, with a responsive and knowledgeable technical support team, is ...

  2. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    Published
    April 13, 2017, 2:30 p.m.
    ...proteins: a relatively pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive GFP variant, super ecliptic...Rosella is a pH-sensitive fluorescent biosensor that was recently deposited with Addgene by Dr. Mark ... and recycling. The key to Rosella’s autophagy-sensing abilities is that its fluorescence emission spectra...regardless of its localization in the cell. SEP’s pH sensitivity means Rosella will fluoresce green unless it’...

  3. It’s Bioelectric! An Exciting Interdisciplinary Field of Research

    Type
    Blog Post
    Published
    Oct. 29, 2019, 2:52 p.m.
    ...channelrhodopsin used as a sensitive fluorescent voltage indicator with near-complete desensitization in continuous... the imbalance of electrical charge between the inside and the outside of the cell. This imbalance is ...Q239 - A GEVI based on the fusion of the voltage sensing domain of a Ciona intestinalis voltage sensor and...anion channel. It is triggered by a short low-intensity blue light pulse and stays open for a longer duration...

  4. Antibodies 101: Conjugation

    Type
    Blog Post
    Published
    March 26, 2024, 1:15 p.m.
    ...spectral properties of the fluorophores you are considering to make sure that they’ll be compatible with ...antibody conjugates (especially if you start to consider clinical applications). But there are a few other... at least mention. Oligonucleotides allow for sensitive detection of low abundance targets with significant...mechanisms. Most take advantage of chemical groups intrinsically present on antibodies, such as the primary amine...can conjugate an antibody yourself allows you to consider a broader selection of antibodies for your experiments...

  5. Developing Lab Management Software for Biology

    Type
    Blog Post
    Published
    Aug. 25, 2016, 2:30 p.m.
    ... learned over the years with the hope that our insights can help steer you in the right direction when... of this is gospel, but we think it's worth consideration. Everything will go wrong Whenever an order ...easily as needed. In this case, the physical considerations of the laboratory make object composition a...technical details of the problem you're solving, but consistency is key. Some namespace collisions you may encounter...treating these pieces of data in ways that are inconsistent with how they are being treated in the lab, ...

  6. 5 Steps to Writing Grant Proposals for Grad Students

    Type
    Blog Post
    Published
    Feb. 11, 2014, 12:13 p.m.
    ...given problem, Empowers the writer, And offers insight into a career as a Professor. Step 1: Your ...sections in a NIH RFA, for example, and tips to consider while writing each section. Research challenge...time to review the literature and offer educated insight into what the desirable features of future technologies...deem your proposal successful. Typical proposals consist of 3 to 5 aims, with secondary milestones to meet... the application. Make sure to invest time and consider the reviewer's comments. Whether the proposal ...

  7. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...virion is a non-enveloped particle of ~100 nm, consisting of a capsid surrounding an inner core which contains...adenovirus: the virion is ~ 100nm in size, and consists of an icosahedral capsid made of 3 types of proteins... for creating adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the ...purification of rAdV uses cesium chloride (CsCl) density gradients combined with ultracentrifugation to ... rAdV genomes/mL) can be measured by: Optical density: particle concentration is measured by looking ...

  8. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 1, 2018, 1:57 p.m.
    ...Interactions Difficult to detect weak and/or transient PPIs. Many PPI identification techniques rely ...often prevents the identification of weak and/or transient interacting proteins. No spatiotemporal control...mass spec analysis. Biotinylated proteins are considered possible interaction partners if they are enriched...BioID, is also useful for identifying weak and/or transient protein-protein interactions since the biotin ...cell lysis and lysate fractionation. Important considerations for Split-BioID It’s important to remember ...

  9. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    ...some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles. Depending on the researcher...ultracentrifugation of the viral stock, followed by resuspension of the collected particles in a smaller volume...Davis et al., 2002). However, polybrene is highly sensitive to freeze-thaw cycles and should be stored in ...enhance transduction efficiency in cells that are sensitive to the toxicity of polybrene (e.g., hematopoietic...PMID: 12050007. 7. Stockschläder, M., et al. "Expansion and fibronectin-enhanced retroviral transduction...

  10. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...III), each responsible for transcribing different types of RNA: Polymerase I is responsible for ribosomal... Rho-independent termination is also known as intrinsic termination, and it relies on the formation of...ribosomal RNA, Polymerase II is responsible for mRNA and miRNAs, and Polymerase III transcribes tRNA and other...polymerase-associated proteins, CPSF and CstF, which are responsible for recruiting the cleavage and polyadenylation...

  11. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    Published
    Aug. 10, 2021, 1:15 p.m.
    ...this temperature and can bind to the primers.  Extension- The temperature is then raised slightly to Taq...  This process of denaturation, annealing and extension is repeated 25-35 times to exponentially replicate...droplet PCR or ddPCR is a method that provides ultrasensitive and absolute nucleotide concentration, unlike...more complicated to develop and is often less sensitive than PCRs that use a single pair of primers, like...Diagnostics NGS library prep QC ddPCR Determine ultrasensitive and absolute nucleotide concentration of starting...

  12. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...because they are inside the virus itself. While these impurities are generally considered innocuous for ...helper plasmids or the cell genome can be packaged inside the vectors during production. These impurities...NGS at Addgene to sequence all the DNA packaged inside the viral particles (Figure 1). Briefly, packaged...contamination is a good start, but to determine the sensitivity of our method we wanted to see how low we could...

  13. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ...injections are most often used when injecting a cell suspension to create xenograft models. Restraint: back scruffing...sounds very disruptive, but surprisingly, it is considered minimally invasive. Restraint: anesthesia Location...common. Due to the intubation, this procedure is considered quite invasive. Restraint: anesthesia Location...introduction to common injection routes in mice! When considering what method you’d like to use, remember that ...animals: routes of administration and factors to consider. PubMed, 50(5), 600–613. https://pubmed.ncbi.nlm.nih.gov...

  14. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...to their high density, which can be collected afterwards. You can then remove the density gradient material...certain in vitro and in vivo applications, you can consider using unpurified AAV from the supernatant (Goodwin...cells and supernatant then need to be purified by density gradient centrifugation (e.g., iodixanol) using...assay for adeno-associated virus vectors with sensitivity sufficient to detect single infectious events...

  15. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

    Type
    Blog Post
    Published
    Dec. 12, 2013, 3:35 p.m.
    ...that onto a mass spec. That's very time intensive and expensive. There's the promise that we could instead...DNA systems capable of storing data and computing inside living cells. He's also a co-founder of the BioBricks...describing the development of “transcriptors,” transistor-like digital genetic switches that enable cellular...proteins. That's central to everything that goes on inside a cell. Until we get a handle on precise, forward...

  16. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...protein combination. The optimized LACE system consisted of: 1) CIBN-dCas9-CIBN, where CIBN is the N-terminal...(Kawano et al., 2015). Nicknamed paCas9-1 and consisting of the fusion proteins N713-pMag and nMagHigh1...able to reach deeper tissues. Additionally, high intensity exposure can cause DNA damage. Far-red light, ...Michael Lin, who developed a chimeric protein consisting of Cas9 and a photo-dissociable dimeric green... anti-CRISPR Acr protein, AcrIIA4, to a photo-sensitive LOV2 domain from A. sativa (Bubeck et al., 2018...editing For more details, check out this comprehensive review comparing optogenetic CRISPR techniques...

  17. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...for your CRISPR genome engineering experiments, consider that wildtype Cas9 with optimized chimeric gRNA...concentration of Cas9 and guide RNA (gRNA). These considerations only matter if the PAM sequence is immediately...to generate off-target DSBs; there needs to be extensive homology of the protospacer followed by PAM. See...design guidelines? A7: A few notes below are considerations for designing HR donor. Generally if off-...away. For small (<50 bp) changes, you should consider using ssDNA oligo as donor template, it's usually...screened? A13: Before starting your experiment, consider co-transfecting with GFP. This allows you to sort...

  18. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    Published
    Sept. 6, 2018, 12:02 p.m.
    ...are some potential complications that must be considered when using this method:     Cells must be amenable...to directly deliver a ribonucleoprotein (RNP), consisting of the Cas9 protein in complex with a targeting...gRNA from a commercial vendor, this can often be expensive. An alternative approach is to express and purify...is most appropriate for a given experiment. Considerations for RNP-mediated CRISPR experiments While using...with these types of experiments. One important consideration is that the Cas9-gRNA RNP will be degraded by...

  19. Transferable Skills Guide: Identifying Your Transferable Skills

    Type
    Blog Post
    Published
    Jan. 11, 2018, 3:29 p.m.
    ... venture presents itself even outside of work, consider challenging yourself, take the opportunity, and... all the things for which you are officially responsible on paper, and take some time to write down all... have to exercise in order to manage these responsibilities. If you are in charge of completing many ...management and organizational skills. If you are responsible for working with lots of students, managing inventories...the short list of transferrable skills here and consider which of these skills you’ve used in your own ...

  20. 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...
Showing: 161 - 180 of 730 results