Skip to main content

We narrowed to 671 results for: FIE

Showing: 121 - 140 of 671 results
  1. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...It was by serendipity that I got into the field of gene therapy, more specifically AAV-based retinal ...trial gone horribly wrong. It brought the whole field to a grinding stop and only now, 2 decades later... work and a little bit of luck A pioneer in the field, Dr. Jean Bennett started working on ways to deliver...Gene therapy was in its infancy, but the eye was identified early on as an ideal target. It is an easily ...have broad tropism in the eye. AAV-2, the first identified serotype and the one used in Luxturna™, can efficiently...
  2. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...studied during graduate school. As a stable, easily purified protein with easily measured activity, MDH is ...Provost’s CURE prepare to analyze their recently purified MDH to study the impact of phosphorylation on ... genuine interest to other researchers in their field. Indeed, sharing results with the broader scientific... original and recombinant enzymes in bacteria, purified them to near homogeneity using a 6XHis tag, and...B. 2020. Mentorship and protégé success in STEM fields. Proceedings of the National Academy of Sciences...
  3. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...has been a long and winding path for many in the field. The vast majority of grant professionals didn’t...and by finding our individual niches within the field of grants. What is a grant professional?  In the...grants has fostered the demand for experts in the field of grants. It is no surprise that the U.S. Bureau...less than 400 international Grant Professionals certified through the Grant Professionals Certification ...by the Grant Professionals Association, and a Certified Social Media Strategist through the National Institute...
  4. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ... Cas9, Cas9 modified 9/12 known off-targets with a 14% indel rate, while ABE7.10 modified only 4/12 off-targets...target base for editing within the editing window specified by the Cas protein These elements were the starting...of dCas9. The resulting editor, BE3, nicks the unmodified DNA strand so that it appears “newly synthesized...efficiency. To further improve specificity, they purified HF-BE3 protein for delivery in ribonucleoprotein... and that they are expressed well. The Liu lab modified the nuclear localization signals and codon usage...
  5. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...that MPG could be engineered to act directly on unmodified G bases, resulting in a G → Y (Y = T or C) base...thymine Since an enzyme that excises inosine can be modified to target guanosine instead, perhaps analogous... et al., 2024). In each case, a UNG enzyme was modified to recognize T and/or C substrates. The HuidaGene...protein engineering going on in the base editing field! From CRISPR-free approaches to mitochondrial base...more diverse. We can’t wait to see what tools the field will come up with next! References and Resources...
  6. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...Many molecular biology techniques require highly purified and concentrated plasmid DNA. This page will discuss...TE. You will now have plasmid DNA that has been purified away from the bacterial proteins and genomic DNA...become sheared and will therefore contaminate your purified plasmid DNA. Incubate solution on ice for 5 min...plasmid DNA. To improve the purity of plasmid DNA purified without a kit it is advisable to perform a phenol...
  7. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...of the primer that binds to the sequence to be amplified (usually 18-21bp). When selecting restriction ... primer that hybridizes to the sequence to be amplified (the ORF in this case), not the Tm of the entire...sufficient amount of DNA). Once you have cut out and purified your insert and vector bands away from the gel...diagnostic restriction digest of 100-300ng of your purified DNA with the enzymes you used for the cloning....
  8. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...imaging system ImageJ or similar software Reagents Purified recombinant antibody, 0.9–1.1 mg/mL SimplyBlue...the recombinant antibody Dilute or concentrate purified recombinant antibody to a concentration of 0.9...platform. Pour off the water in the sink. Take a brightfield image of the gel with an appropriate imaging ... that have a purity of ≥90% but some labs use unpurified tissue culture supernatant routinely without ...
  9. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...isolation of modified cells; and (5) is compatible with the introduction of unique molecular identifiers, which... In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However... of the pRDA_526 vector. Moreover, the screen identified putative novel regulators of CD8+ T cell responses...
  10. Finding Your Science Policy Path

    Type
    Blog Post
    ...many science policy professionals that entered the field from a more traditional policy background, learning...groups, attend webinars, and learn more about the field.  Read the CCST Science Policy Career Guide to learn...   Where can you do science policy? Because the field of science policy is so diverse, you can do it in...once selected, you can pivot to a completely new field without issue. This may be more challenging if you...you are trying to apply for an outright job in a field where you have no specific scientific credentials...
  11. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ... editing. Repair oligos do not need to be PAGE purified, although doing so increases knock-in efficiency...CRISPR in C. elegans The Meyer and Mello labs also identified several experimental set ups that could improve...choosing. Tools such as SapTrap Builder greatly simplifies design of these constructs, allowing novice researchers...Developments in mammalian and yeast cells using modified Cas9 proteins to regulate gene expression (CRISPRi...systems, allowing rapid progress in a wide range of fields.   Thank you to our guest blogger! Jordan Ward...
  12. AAV ddPCR Titration

    Type
    Protocol
    ...protocol to perform digital droplet PCR (ddPCR) for purified AAV titration....how to use droplet digital PCR (ddPCR) to titer purified recombinant Adeno-associated viral vectors (AAV...the ITR elements in the viral vectors but can be modified for other targets. The dilution series outlined...ddPCR for AAV quantification. This protocol was modified from the publication Lock et al. (2014) (Link ...
  13. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    ... process, the insert and vector are first PCR amplified. The forward primer used to amplify the vector...PCR, you should confirm that the products were amplified by running a small aliquot of the reaction on ...2. Li, Chaokun, et al. "FastCloning: a highly simplified, purification-free, sequence-and ligation-independent... Our Special Collections for Plasmids from Your Field ...
  14. Cellosaurus: A Cell Line Information Database

    Type
    Blog Post
    ... cell lines used in scientific research are misidentified! Rigor and reproducibility of scientific data...and collections. Importantly, Cellosaurus also identifies problematic cell lines: ones either suspected...or easily contaminated, as well as commonly misidentified lines. Browsing this feature to know if your... care about. If you have cell lines without a certified origin (you know, that split you got from the ...
  15. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...technology to survive the diverse demands of the biotech field, let's look at how prokaryotes originally utilized...generate RNA, which guide a Cas nuclease to the specified exogenous genetic material next to the species-specific...re-identifying it (3). Surprisingly, few genes have been identified that neutralize CRISPR/Cas. Earlier this year...lab protocols, providing tips from experts in the field, and enabling access to multiple plasmids used for...
  16. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ... region. The variable region acts as a unique identifier for individual cells. Each cell is labeled with.... The fixed region acts as a library-specific identifier and allows cells to be tagged with different ...during reprogramming. These library-specific identifiers act like a timestamp and help demultiplex the...this lineage tracing experiment, the Morris lab identified a novel gene, Mettl7a1, whose expression helped...
  17. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ...small molecules since the very beginnings of the field. Those engineered sensors were designed to replace...1980s. “Synthetic dyes were the standard in the field, but there is one problem: how to get that into ... sensors Griesbeck’s recent contribution to the field is a series of fluorescence resonance energy transfer...protein, troponin C (TnC), from muscle cells. They identified a toadfish TnC variant which had a high-affinity...
  18. New and Upcoming Viral Vectors - June 2019

    Type
    Blog Post
    ...pAAV-CAG-GFP 59462  AAV2 pAAV-CAG-tdTomato (codon diversified) 114472  AAV5, AAV8 pAAV-hSyn-mCherry 50459...for tracking large populations of neurons in wide-field imaging (Dana et al., 2018). See our full biosensor... For an estimate on availability date or to be notified when a particular catalog item is available for... the vector name and serotype you’d like to be notified about. Controls Plasmid Serotype Name 37825...
  19. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    ...selectable marker, and target gene, for example, are amplified with overlapping regions of homology and inserted...for transformation into yeast that same day.  The field of synthetic biology has led to advances in biofuel... in these methodologies offer streamlined and simplified alternatives to traditional cloning. NEBuilder...and opens the door for new opportunities in the field.  Gibson Assembly® is a registered trademark of ...
  20. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ...conformational changes. These tiny movements are then amplified tens or hundreds of thousands of times by propagating...membranes outside of their natural context or 3) be modified to extend at different pH levels. Domesticating...soluble when extended (see figure to the right), we identified R-body mutants that extend at both lower and ...response to pH changes. Beyond these applications, purified R bodies could also perform as actuators in microelectromechanical...
Showing: 121 - 140 of 671 results