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Showing: 321 - 340 of 705 results
  1. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    Published
    Dec. 15, 2017, 2:07 p.m.
    ...to neuroscience prior to this conference was a rotation in Qiao Zhou’s lab in graduate school where I ...neuroscience and you don’t have a background in statistics and programming, you’d be wise to start working...and brain monitors. Walking around the poster presentation area, I honed in on projects using biological...
  2. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    Published
    May 9, 2017, 2:12 p.m.
    ... efficient. Pro-Tip Make sure to reverse the orientation of the crRNA’s direct repeats when expressing...preventing packaging of the virus.  Reversing the orientation of the direct repeat protects the (+) stranded...cells, but notice how Zetsche et al reverses the orientation of the promoter driving expression of the crRNA...
  3. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    Published
    May 26, 2020, 1:15 p.m.
    ...this one mutated methionine was not enough to prevent alternative initiation, so they mutated the methionines...the target site (the same sequence you’d like to mutate in the genome) and an out-of-frame H2B (histone...
  4. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ...long sequences with reduced risks of unwanted mutations in mammalian cells. Long sequences traditionally...traditionally have higher risks of off-target mutations. These plasmids contain catalytically inactive dCas9 and...challenge due to multiple factors, including limitations of number of flourescent proteins that can be...
  5. Antibodies 101: ChIP

    Type
    Blog Post
    Published
    March 10, 2022, 3:25 p.m.
    ...Chromatin immunoprecipitation (ChIP) is an extremely useful technique that provides insight into protein...to counteract the effects of increased sample agitation; a shorter sonication time could be an opportunity...resources on Addgene’s blog Antibodies 101: Immunoprecipitation  Antibodies 101: Polyclonal Antibodies Antibodies...
  6. Hot Plasmids - February 2022

    Type
    Blog Post
    Published
    Feb. 22, 2022, 2:15 p.m.
    ...Cas12a for multiplexed gene perturbation The Penn State Protein Ladder Engineered miniature CRISPR-Cas system...2021. doi: 10.1038/s41587-020-0600-6. The Penn State Protein Ladder  by: Rachel Leeson This simple, inexpensive...expression and purification. Figure 3: The Penn State Ladder run on SDS-Page gels. A: individually expressed...
  7. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    Published
    Jan. 25, 2024, 2:15 p.m.
    ...there is a classification system, CasID, to help facilitate direct comparison of different enzymes. CasPEDIA... their sequence (NGG, etc.) as well as their orientation relative to the gRNA (5’ or 3’). In some cases...would be helpful to know for viral packaging limitations. Maybe you want to read literature on the less...
  8. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    .... While certain cell types, such as principal excitatory cells and glia, have been successfully targeted...researcher’s locus of interest. NHEJ- or HDR-induced mutations in ATP1A1 can render cells resistant to the small...improves editing efficiency. Importantly, the point mutations created by Agudelo et al. prevent ouabain binding...DNA molecular weight markers The Tan lab at Penn State - set out to “rule” Molecular Biology. Under the...
  9. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    ...than indirect ones and can be used in highly quantitative assays. On the other hand, they are not as sensitive...stability Limited protein size range Tris-acetate 40–500 kDa Tris-tricine 150 V, 1–3 hours ...detection. Film also needs to be digitized for quantitative work — and with the advent of online lab notebooks...high-abundance single protein controls in semi-quantitative immunoblotting. Journal of Neuroscience Methods...
  10. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    Published
    June 27, 2019, 12:25 p.m.
    ...altering gut microbiome members in their native habitat (Ronda et al., 2019). They called their method ...the addition of standards, the assay is semi-quantitative and can approximate mRNA concentration within...infection, dysbiosis, and antibiotic treatment. To facilitate these studies, Pamela Silver’s lab developed ...
  11. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ... control the abundance of proteins in cells. Homeostatic regulation of protein abundance is essential ...Alternatively, fluorescent protein tags can be used to facilitate the visualization of specific proteins in cells...proteins in situ. Now FUNCAT-PLA provides both quantitative and spatio-temporal information about the turnover...
  12. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    Published
    July 25, 2017, 1:20 p.m.
    ... pathway of interest, but there can be a lot of static since other signaling networks can also be non-....” Dronpa’s color thus indicates MEK1’s current state (Green when off, dark when on). Figure 1: Photoswitchable...turn off psRaf1). After this, the phosphorylation status of MEK1/2 was tracked at 5 minute intervals via...
  13. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    Published
    June 15, 2017, 2:30 p.m.
    ...complicating the interpretation. Because of this, these methods only give a qualitative view of dimerization...molecules based on size is ultracentrifugation. Sedimentation equilibrium analytical ultracentrifugation yields...
  14. Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages

    Type
    Blog Post
    Published
    Jan. 11, 2017, 3:30 p.m.
    ...for compliance. However, 10 days prior to the implementation date, our data indicated that, while 69% of...the lapsed mandate (or are being compelled at the state level, in the case of some public institutions) ...continuing to reach out to institutions with a statement indicating that we encourage them to keep raising...
  15. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    Published
    March 15, 2022, 1:15 p.m.
    ...assay for a native (folded) protein, such as immunoprecipitation, the epitope may not be accessible for binding... down in a variety of ways. First, review the citations. An antibody that has been cited thousands of ...antibody to that of the anti-tag antibody; and immunoprecipitation-mass spectrometry strategies that isolate...
  16. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ...vector with half the size of the ssAAV genome has a mutation in the terminal resolution site (TRS) to form ...genome with wild-type ITRs at the both ends and mutated ITR at the center of symmetry. After uncoating ..."Development and optimization of a real-time quantitative PCR-based method for the titration of AAV-2 ...
  17. Addgene: The First Twenty Years

    Type
    Blog Post
    Published
    Dec. 19, 2024, 2:15 p.m.
    ...to start an open science nonprofit that would facilitate the sharing of plasmids between scientists. The... r/labrats, Reddit.  Part of that work meant facilitating the Material Transfer Agreement process, which... millionth item, and had a cumulative 120,000 citations of individual items in our repository, a tribute...
  18. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    Published
    Jan. 14, 2016, 3:30 p.m.
    ...plasmid are the backbone, inserted gene/functional mutations, antibiotic resistance, restriction sites used...most sequence assembly errors occur), deletions, mutations, in-frame protein fusions, and multiple cloning...cases, we deal with multiple inserts or internal mutations that require more than two sequencing reactions...
  19. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    Published
    Jan. 29, 2015, 1:25 p.m.
    ...A professional coach will provide an initial consultation meeting or call for free.  Take advantage of...affordable for you. There is also a group The Dissertation Coach based in Illinois. A few of the coaches...in this group have scientific backgrounds. For a stated purpose of transitioning a scientist from academia...
  20. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ...conditions (individually versus in groups), health status, diet, cage enrichment, stress level, origin (vendor...of channel protein, they can trigger neuronal excitation or inhibition upon light (or drug) application...situations such as learning and memory or emotional states, for example anxiety or depression. In upcoming...
Showing: 321 - 340 of 705 results