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We narrowed to 845 results for: CAL;

Showing: 541 - 570 of 845 results

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

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...on DNA, or are enzymatically active. For this reason, recombinant proteins are typically expressed in E...medium. Induction timing is important, as you typically want to make sure your cells have first reached...your expression conditions by setting up a small-scale expression experiment to test variables such as ...

  2. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...control over where mutations are introduced, and scalability (see Supplementary Table 1 of Wrenbeck et al)...stretches of sequence could be used. However, a practical limitation is sequencing depth and cost. You need...Measuring the Activity of Protein Variants on a Large Scale Using Deep Mutational Scanning.” Nature Protocols...scanning: assessing protein function on a massive scale." Trends in Biotechnology 29.9 (2011): 435-42. PubMed...

  3. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ...ICC or a tissue in the case of IHC. IF is used specifically when we visualize the antibodies using fluorescence...counterstaining, mounting sample on slides, and confocal   Figure 2: High-level overview of an ...controls to show that your antibodies are binding specifically. For positive controls, aim to use samples that...in an organism that is underrepresented in the biomedical research community. You may have to take some...

  4. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    Published
    March 1, 2016, 3:30 p.m.
    ... regions of homology at their ends, which are typically created by PCR. Then, the fragments are incubated...competent cells. The success rate of this reaction is typically fairly high, so there is usually no need to screen...the bacterial chromosome (up to 100 kb) were specifically cut out via CRISPR and then assembled into a...amplify the fragments of the chromosome, which was technically challenging. Other homology based technologies...

  5. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    Published
    April 5, 2018, 12:42 p.m.
    ...simple in C. elegans and bacteria. But CRISPRing typical mouse models, including creating Cre-dependent ...on both sides and the T7 promoter on one end. Typically, Easi-CRISPR works best with insertions of 2 kb...models varies from 8.5-100% of pups born, with typical efficiencies of 30-60%. Quadros et al. attribute...2.5x more animals than i-GONAD and is a more technically difficult process. Overall, the main drawback...

  6. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Sept. 24, 2020, 1:15 p.m.
    ...desired location of edit; efficiency decreases dramatically when the cut site is >30nt from the proximal... constraints are even more exquisite for the so-called base editor Cas9, which makes DNA changes in the...nuclease-mediated off-target effects.” Further, large-scale datasets of hundreds of genetic screens using genome-wide... sequences targeting a gene to score in order to call that gene a hit.  This is an exciting time for functional...

  7. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...facilitates the integration of the transposon specifically at ‘TTAA’ sites randomly dispersed in the genome...re-integration from the piggyBac™ donor, which is typically found with the wild-type piggyBac™ transposase..., including making transgenic cell lines and genetically-modified cell lines. Furthermore, the use of ...cell engineering team.     Mathew Goodwin is a Technical Sales Specialist at Transposagen. As a gene editing...

  9. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    Published
    June 4, 2024, 1:15 p.m.
    ... BioRender.com.   Cas9 has two major lobes, called NUC (for nuclease) and REC (for recognition). The...recognizing the PAM on DNA, the CTD is also sometimes called the PAM-interacting (PI) domain. The guide RNA ...conformational state of the HNH domain also allosterically controls RuvC activity through a hinge region...Visualization, and Informatics at the University of California, San Francisco, with support from National Institutes...

  10. 5 Science Rap Videos & Their Creator: Tom McFadden

    Type
    Blog Post
    Published
    Feb. 9, 2017, 3:30 p.m.
    ...African American woman whose cervical cancer cells became a mainstain of biological study without her or her...fossil rocks and their importance in understanding biological history. 4. Please Don't Kill My Hive Beautifully...

  11. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    Published
    May 2, 2024, 1:15 p.m.
    ...important immune cell type, in more complex and physiological microenvironments. These screens have primarily...molecular identifiers, which can improve the statistical power of in vivo screens. Figure 1: Using...Fellow in Arlene Sharpe’s laboratory at Harvard Medical School and is interested in improving CD8+ T cell...

  12. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...multiple timepoints. When the target of interest is critical to cell survival or proliferation, an additional...a flow cytometry experiment. But validation is critical to ensuring accurate interpretation of your results...to start with a screening procedure in easily genetically modified cell types (top). The specific approach..., A. M., McPherson, P., & Laflamme, C. (2023). Scaling of an antibody validation procedure enables quantification...

  13. Bringing Sustainable Practices to the Lab: Easy Action Items

    Type
    Blog Post
    Published
    April 22, 2019, 12:26 p.m.
    ...alternatives for some of the harmful chemicals in your lab. These chemicals are not only better for the environment... resources about this, and even has a list of biological samples that are stored long term at -70 °C. ...

  14. What's New in CRISPR - Winter 2018

    Type
    Blog Post
    Published
    Dec. 18, 2018, 1:17 p.m.
    ...plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools more ... Feng Zhang’s lab recently developed a platform called SHERLOCK (specific high-sensitivity enzymatic reporter...will help researchers map transcription factor localization and binding via ChiP-seq. Learn more about...

  15. Make a Splash: Notions of Scientific Impact Are Evolving

    Type
    Blog Post
    Published
    Dec. 5, 2014, 2:54 p.m.
    ...papers detailing Nobel Prize-winning research typically weren’t among the most cited. And, as scientific...online, surely times have changed. Altmetrics.org calls to expand our view of what impact looks like and... aren’t enough. Scientists are sharing what they call “raw science” – data sets, code, and, I think it...

  16. A Scientist's Guide to Ebola

    Type
    Blog Post
    Published
    Oct. 16, 2014, 3:01 p.m.
    ...protein present on the virion surface – making it critical for viral attachment and fusion. Once inside a...glycoprotein’s role in infection also makes it a critical component of vaccines and the target of neutralizing...established Ebola Genome Portal at the University of California, Santa Cruz, offers sequence data on five strains...

  17. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    Published
    Jan. 23, 2014, 3:23 p.m.
    ...few years ago, researchers introduced what’s been called a major breakthrough in cloning technology with...cassettes onto a single T-DNA in subsequent rounds, dramatically reducing the time required to generate multi-construct...multi-construct transgenic plants. In the most practical terms, what GreenGate can do is “save you half a...

  18. Top Requested AAV of 2017: pmSyn1-EBFP-CRE

    Type
    Blog Post
    Published
    Jan. 17, 2018, 2:57 p.m.
    ...promoter. It can be used to drive Cre expression specifically in neurons. AAV Delivered Cre - A useful tool...Indeed, this AAV provides you with a means to specifically activate expression of any Cre-dependent gene...Additionally, you could use the retrograde form to specifically express Cre in neurons with axons that extend...

  19. Editor's Choice, August 2016

    Type
    Blog Post
    Published
    Sept. 2, 2016, 1:32 p.m.
    ...sequence in live cells. This technique uses catalytically inactive CRISPR endonuclease (dCas9) fused to...scientists. You can find all of our August posts in chronological order here: Site Directed Mutagenesis by PCR...Single Base Editing with CRISPR Cas9 Activators: A Practical Guide 10 Steps to a Perfect Science Talk Developing...

  20. Scientist Networking: What is an Informational Interview?

    Type
    Blog Post
    Published
    July 1, 2014, 3:42 p.m.
    ...meet interesting people doing interesting things. I call this building relationships because networking has...or a particular position at their company, do NOT call this an Informational Interview. If you do this,... if it is not possible consider a phone or video call. If you do decide to use Skype, make sure to follow...

  21. Top 10 Open Science Developments of 2013

    Type
    Blog Post
    Published
    Dec. 19, 2013, 3:46 p.m.
    ...Science and Technology Policy issued a memorandum calling on Federal agencies with over $100 million in annual...promote open science, ensure good stewardship of clinical trial data, serve society and patients, and respect...patents in a case against Myriad, an organization called Free the Data launched a public database to gather...

  22. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    Published
    Aug. 4, 2015, 2:30 p.m.
    ...used a GFP derivative to conduct subcellular localization studies or make a reporter construct. Fluorescent... mammalian cells, and it may find wide use in biological research. Photoconvertible fluorescent proteins... new transgenic line could avoid problems in localization that have been encountered by some investigators...

  23. Easing the Protein Purification Process with pCri

    Type
    Blog Post
    Published
    June 19, 2015, 3:08 p.m.
    ...research, notably determining the subcellular localization and the number of disulfide bonds in your protein...protein expression in systems other than E. coli, specifically the bacterium Bacillus subtilis and the yeast...may improve expression of proteins that natively localize there. pCri is both versatile and easy to use,...

  24. What's New in CRISPR - March 2020

    Type
    Blog Post
    Published
    March 10, 2020, 1:15 p.m.
    ...plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools more ...and an optogenetic tool that controls nuclear localization (LINuS). Blue light renders the NLS accessible...expression of specific Caulobacter genes using a catalytically dead Cas9 (dCas9) derived from Streptococcus...

  25. Supporting Rare Disease Research with BeHeard 2017

    Type
    Blog Post
    Published
    July 20, 2017, 2:30 p.m.
    ...Institute partners with companies to provide technological resources for those studying rare disease. This...will use fluorescent proteins to monitor the localization of these genes, CRISPR to disrupt their function... University of British Columbia Department of Urological Sciences. Dr. Nabavi studies peritoneal mesothelioma...

  26. Working 9-5: an Addgene Co-op

    Type
    Blog Post
    Published
    June 2, 2022, 2:15 p.m.
    ...hour long interview. I anxiously prepared for the call. The length of this interview was intimidating to...team and shows how they fit into the system. Specifically as a co-op, it is encouraged to explore different... a 9-5 As it turned out, Addgene uses something called “core hours”. Core hours consist of the time between...

  27. Addgene is Expanding Our Viral Vector Service!

    Type
    Blog Post
    Published
    Oct. 15, 2024, 1:15 p.m.
    ...service. A small subset of AAV plasmids will have technical or legal restrictions that make them ineligible...be reviewed by Addgene for legal approvals and technical compatibility with our production process. This...found in our in-stock preps. If a request has technical considerations that may affect expected titers...

  28. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

    Type
    Blog Post
    Published
    Aug. 13, 2014, 3:49 p.m.
    ...parts. Their efforts yielded a protein, which they call SpyLigase (Spy comes from the “S” in Streptococcus...able to grab on to proteins irreversibly in a genetically encoded way. SpyTag is nice and small, just 13...this tool? Howarth: Polyproteins are useful for mechanical study of proteins by atomic force microscopy ...

  29. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    Published
    July 29, 2014, 6:16 p.m.
    ...the collection without getting overwhelmed by its scale: Look at the mEmerald variants first, followed by...fluorescently tagged versions of many proteins known to localize to specific cellular structures.The Davidson lab...you’re not sure where your protein of interest localizes in the cell, use these markers as your guide. ...

  30. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    Published
    June 11, 2020, 1:34 p.m.
    ...wavelength of light. This response is immediate, on the scale of milliseconds, and can precisely link a neuron's...control over timing. Because chemogenetics relies on chemical activation and the diffusion of drugs throughout...require a permanent intracranial implant to be surgically installed to allow for light delivery. Chemogenetics...
Showing: 541 - 570 of 845 results