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Showing: 341 - 360 of 1012 results
  1. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ...where droplet digital PCR (ddPCR) comes in. AAV titering using quantitative PCR Before we dive into the... PCR is complete, the results are analyzed by comparing the threshold fluorescence of the viral DNA to...reasons, we are in the process of transitioning our titering method from qPCR to ddPCR. Droplet digital PCR...reference, meaning saving on reagents (and time)! AAV titering using droplet digital PCR Droplet digital PCR ...generator, you will have to be careful about transferring your droplets from the droplet generator to ...detection of microbes. Some applications include measuring viable probiotics and detecting circulating pathogens...
  2. SciComm: Taking Science to Elementary and Middle Schools

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    Blog Post
    ... parents in STEM careers, chances are they’re wondering how they could contribute to the school, too. ...Middle school: supplementing the science curriculum Sharing my expertise with our district middle school was... an authority in this area. What is genome engineering? How does CRISPR work? This section was essentially... of the talk by running it by the teachers and sharing some slides on Twitter, where I was able to get...computers and the internet before it, genome engineering technology is moving along much faster than it...understanding genetics is so timely.” Genome engineering, and many other technologies and fields in biology...
  3. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

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    Blog Post
    ...the non-radiative transfer for energy between neighboring fluorophores, FRET can be used to detect the ...ratios) to generate useable data. What’s more, measuring and quantifying FRET is often easier said than...complete FP because the PIPs they’re tethered to bring them into close proximity (Figure 1). Formation ...independent of PPIs. Overexpression can be assessed by comparing the expression levels between BiFC constructs ...Analysis of Protein Interactions in Live Cells. Cold Spring Harb Protoc 2013:pdb.prot076497. https://doi.org...Fluorescent Protein 101 blog posts Read our blog post covering an introduction to FRET Resources on Addgene....
  4. Hot Plasmids: Fall 2024

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    Blog Post
    ...But CRBN has been difficult to study in vitro, requiring either expressing it with additional cofactors...faster, with expression levels comparable to GFP during early embryogenesis (Figure 3). mScarlet-I3 also... through the middle of each embryo are shown, comparing GFP, wrmScarlet (mScarlet), and the new mScarlet3...proteins of interest) and mScarlet-I3 for scenarios requiring rapid maturation and prolonged imaging with minimal...high rate of spontaneous antibiotic resistance interfering with selection. Using an inducible Cas9 expression...to cause disruption of the target gene. Lastly, curing the mCherry-sgRNA plasmid leaves an edited strain...
  5. CRISPR Activation: A Practical Guide

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    Blog Post
    ...the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 is an enormously plastic tool and...activator by simply providing a new gRNA rather than engineering an entirely new protein. This makes it much cheaper...types of Cas9 activators, check out our blog post covering VP64, Suntag, SAM, and VPR. Worry less about off...the Wyss Institute for Biologically Inspired Engineering.     Alejandro (Alex) Chavez is a member of the...develop several popular CRISPR-based tools for altering DNA sequence or regulating gene expression.   ...Church GM (2015) Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach....
  6. Save Time with Transient Plant Leaf Transformations

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    Blog Post
    ...are syringe or vacuum infiltration. Syringe infiltration (D'Aoust et al., 2009): Usually a syringe without...contains a plasmid with a T-DNA (transfer-DNA) region. During infection this T-DNA region is transferred into...transformation and gene expression can be tracked. During a transient leaf transformation, the engineered...Transient expression of antibodies in plants using syringe agroinfiltration." Recombinant proteins from plants...
  7. Designing Your Chalk Talk for the Academic Job Interview

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    Blog Post
    ... You need to balance respectfully answering questions while steering the conversation back to your vision...proposal or a 10-year research plan. I suggest preparing for both types. Your research vision is your approach...proposal will distinguish between competing models. Comparing model A vs. B is more constructive than testing...vision. On the day of your chalk talk Bring your own fresh set of erasable markers. You’ll have the colors...and consider eating one before your chalk talk. Bring your own bottle of water. If possible, write down...
  8. To Codon Optimize or Not: That is the Question

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    Blog Post
    ...optimization? Codon optimization is an approach in gene engineering to improve gene expression by changing synonymous...efficiency and thus protein expression without altering the sequence of the protein. As DNA synthesis ...tRNAs, including those that exhibit wobble base-pairing (a tRNA that can recognize multiple synonymous ...of translation. The ribosome may slow and pause during elongation which may actually be necessary for ... SA, Zhou W, Mauro VP (2005) An mRNA-rRNA base-pairing mechanism for translation initiation in eukaryotes...02)00362-x Stadler M, Fire A (2011) Wobble base-pairing slows in vivo translation elongation in metazoans...
  9. Overcoming the AAV Size Limitation for CRISPR Delivery

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    Blog Post
    ...inteins. Split inteins are a pair of naturally occuring polypeptides that, when at the ends of two proteins...Marraffini LA, Zhang F (2013) Multiplex Genome Engineering Using CRISPR/Cas Systems. Science 339:819–823...Spinner HB, Baney KLM, Chuck J, Tan D, Knott GJ, Harrington LB, Al-Shayeb B, Wagner A, Brötzmann J, Staahl..., Grimm D (2014) CRISPR/Cas9-mediated genome engineering: An adeno-associated viral (AAV) vector toolbox...Gao L, Makarova KS, Koonin EV, Zhang F (2019) Engineering of CRISPR-Cas12b for human genome editing. Nat... Mary Gearing...
  10. Replacing Paper: Tips for Choosing an Electronic lab Notebook

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    Blog Post
    ...briefing, to quick notes, to finances, to flights & ordering food - everything is covered.   So why are we ...folders in complex ways, there are solutions that bring more to the table in terms of data traceability ...researchers today generate vast amounts of digital data during their work. It is already becoming impossible to...to keep all their data in one place and avoid scattering intellectual property across various software...functionalities include comments, notifications, sharing files, assigning team members to certain tasks,...compliance requirements – e.g. 21 CFR part 11, HIPAA? Answering this question helps you evaluate whether the ELN...
  11. Hot Plasmids: February 2026

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    Blog Post
    ...through wobble pairing (Figure 4). Now, through a massive effort of protein and genome engineering, the Rinehart...like heat and oxidation or protocols for tissue clearing and correlative light and EM imaging. ... acids into reporter proteins. This feat of bioengineering could allow fast and cheap production of synthetic...bacterial strain here! Grome, M. W., et al. (2025). Engineering a genomically recoded organism with one stop ...Doench and David Root have created the next expert pairing of Jacquere and Julianna, lentiviral CRISPR knockout...
  12. 10 Steps to a Perfect Science Talk

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    Blog Post
    ...about the particular audience and venue before tailoring each presentation appropriately.  Find out who...many non-scientists be present?  Always ask the hiring manager or host for topic advice. An added bonus...figure out how to make it suspenseful. 4. Start preparing your talk early - like today Don’t wait until ... all graduate students and postdocs form peer mentoring groups to practice giving talks and getting feedback...always have a few extra slides in their back pocket. During your practice rounds, encourage your listeners ...down it is a great segue to talk about how the sharing model works. See if you can elicit helpful comments...
  13. Antibodies 101: Introduction to Gating in Flow Cytometry

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    Blog Post
    ...interested in measuring a population based on a fluorescent stain, you might be wondering which second ...types of scattering that occur when a cell is pulsed by the laser. Within each type of scattering, you also...to keep it short though, promise! FSC and SSC: scattering To digitally extract your cells based on their...cells stain positively for both markers. When comparing two markers simultaneously, we commonly describe...
  14. Editor's Choice, October 2016

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    Blog Post
    ...important information for anyone considering a postdoc. In “Changing Labor Laws Bring Increased Postdoc Wages,...readership with over 75,000 views. Changing Labor Laws Bring Increased Postdoc Wages Our most popular blog post...Grad School Advice Part 2: Building Community Engineering the Plant Genome Using CRISPR/Cas9 Healthcare...Reddit for Science Communication Changing Labor Laws Bring Increased Postdoc Wages I hope you all have a great...
  15. 2022 and 2023: Reflections and Looking Forward

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    Blog Post
    ... what challenges remain in materials sharing, information sharing, and research reproducibility while ...service and support.  Alongside our material offerings, in 2022 we also rolled out an updated AAV Data...around the world can upload AAV validation data, sharing what works and, equally importantly, what doesn...commitment to supporting the research community through sharing science and distributing high-quality materials...
  16. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

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    Blog Post
    ...relatively simple in C. elegans and bacteria. But CRISPRing typical mouse models, including creating Cre-dependent...Megamer service tested by Quadros et al. When delivering the ssDNA template and CRISPR machinery, Quadros...facilities for 13 projects, with most projects requiring only 40-60 zygotes to generate a correctly edited...conditional alleles by CRISPR/Cas-mediated genome engineering." Cell 154 (2013):1370-9. DOI: 10.1016/j.cell... a robust method for in situ germline genome engineering using CRISPR nucleases. Genome Biol. 19 (2018... Mary Gearing...
  17. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ...Westerns are divided into two categories, native and denaturing. In a native western, the protein’s secondary...is separated through a matrix by charge. In a denaturing western, the protein is denatured to its primary... purpose of this blog, we will be focusing on denaturing westerns.  Separating a protein mix by size Sodium...The wet transfer process is very effective in transferring a broad range of protein sizes but takes hours...blocking the membrane  from binding to the antibodies during the stain. The ideal blocking buffer depends on...loading control when assessing protein abundance or comparing samples across lanes.  For a typical loading control...
  18. Download Addgene’s Science Career Guide, Plus 8 Career Tips from Addgenies

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    Blog Post
    ...social media, you could be sharing your research or finding opportunities to bring science to the community...the same page with the potential PI in terms of mentoring style and expectations. Make sure you will enjoy...mantra and never forgot it. Usually when I see inspiring artists who are thinking about getting into the... the process down into steps and set aside time during the week to tackle each one.  Get the toughest ...
  19. IDP and your PI: A Roadmap for Career Planning and Personal Development

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    Blog Post
    ...year, graduate students and post-docs alike are wondering where the time has gone. Are we any closer to ...to graduating, publishing that key paper, or figuring out a career path? Many trainees are developing Individual...accomplishments, from learning new techniques to finally figuring out a tough experiment, sets a positive and encouraging...Good Manager Anyway Developing Transferable Skills During Science Training ... Mary Gearing...
  20. Tag Your Favorite Yeast Genes with Ease

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    Blog Post
    .... pombe. The protocol is surprisingly simple, requiring only a PCR product containing the modifying sequence...affinity purification of protein complexes. John Pringle and Jürg Bähler have deposited a large collection...genome modification in yeast developed by Dr. Pringle’s former lab at UNC Chapel Hill. Bähler et al.(2...McKenzie A 3rd, Steever AB, Wach A, Philippsen P, Pringle JR. Yeast. 1998 Jul;14(10):943-51. Longtine MS,... DJ, Shah NG, Wach A, Brachat A, Philippsen P, Pringle JR. Yeast. 1998 Jul;14(10):953-61. Note: Marcy...
Showing: 341 - 360 of 1012 results