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Showing: 321 - 340 of 1016 results
  1. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

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    Blog Post
    ...vector into the culture medium during adeno-associated virus manufacturing." Human gene therapy21.10 (2010...Enhancement in vector stability." Biotechnology and bioengineering 94.2 (2006): 322-329. PubMed PMID: 16528756...pressure and sodium butyrate." Biotechnology and bioengineering 42.5 (1993): 601-610. PubMed PMID: 18613082...and antibody production." Biotechnology and bioengineering 37.10 (1991): 989-993. PubMed PMID: 18597326... Kamen. "Hyperosmotic pressure on HEK 293 cells during the growth phase, but not the production phase,...
  2. Finding Your Science Policy Path

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    Blog Post
    ...ranges from astronomy to bioethics to chemical engineering and everything in between. Plus, there are many...pursue it. It can manifest via research and data gathering, regulatory efforts, policy implementation, program...societies host “Hill Days” where they train and bring researchers to Congress to advocate for increased...normal!  Action Items:  Start a career journal during your current job and note what makes your heart...writing and presenting for non-scientist audiences during your time in graduate school through outreach organizations...working in student government, and teaching and mentoring others (see more in Figure 2). Starting early ...programs can connect you with alumni before and during your application process so you can learn more!...
  3. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

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    Blog Post
    ...reagent-grade endangered Martian chicory root, but considering this is a needle in a haystack type of objective...In retrospect I used a way too high Cas9 MOI considering that A549 cells are relatively easy to transduce...could avoid potential off target artifacts by comparing the phenotypes of multiple gRNAs. Targeting different...lanes 0.3 % 3.0, Figure 5, lane 2) is faint, so comparing it to the undigested control (Figure 4, lane 0...straightforward, and is a testament to how far genome engineering has come. For comparison, in 2007 it took about...for Designing Primers UCSC Genome Browser: For eploring genomic DNA ImageJ: Free Software for Analyzing... genome engineering...
  4. Technologies Enabled by NanoLuc® Luciferase

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    Blog Post
    ... Kyle Hooper at Promega. Researchers have been sharing plasmids ever since there were plasmids to share...applications of the Enhanced Nano-Lanterns including monitoring multiple cellular events such as the dynamics...protein binds to calcium it changes conformation and brings the two fluorescent proteins together thereby increasing... blue-shifted fluorescent protein (donor) and measuring the resultant emission from the red-shifted fluorescent...release with an optogenetic receptor, melanopsin, requiring brief exposure to 470nm blue light. To avoid overlap...is available through Addgene (Plasmid # 87127). Pairing NanoLuc® luciferase with fluorescent proteins has...in vivo optical imaging and flow cytometry for monitoring tumorigenesis. Cancer Res. 75, 5023-33. PubMed...
  5. Visualizing Translation at the Single Molecule Level

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    Blog Post
    ...translation is key to cellular function, especially during development or stress. With ribosome profiling,...to track translation in two different ways: by monitoring the first round of translation or by tracking...NLS-MS2-RFP) the reporter mRNA, which contains the tethering sites for the fluorescent proteins and is driven...double labeled, indicating translational repression. During later stages, single RFP-labeled osk and Oskar ...with the red reporter mRNA. Yan et al. used harringtonine, a translation inhibitor that stalls ribosomes... Mary Gearing...
  6. Summer SciComm Series: A PhD in Science Communication

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    Blog Post
    ...how it works. I used to stay up past my bedtime, poring over my grandparents’ medical textbooks by torchlight... I asked participants to read short abstracts comparing the mortality rates between two controversial ...and biased. As a result, the control abstract comparing two fictional medical issues – chronic and acute... Kahan et al., 2012). Scicomm in the community During my PhD, I’ve also found opportunities for more ...community. I brought along printed copies of studies comparing the filtration efficacy of different materials... in technical jargon that even scientists in neighbouring disciplines might struggle to understand. Why...
  7. AAV Q&A with Tim Miles

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    Blog Post
    ...to keep their cargo below 4.4kB (with the ITRs bringing total genome length to ~4.7kB.  If your insert... kB, then you would be oversized even before considering the need for some manner of polyA. In this scenario...intein approach.   Q3: What’s the best method for titering AAV? Is there a difference in titer results between...equal animal dosing, we advise that users keep titering method consistent across samples within a project...microglia in vivo.   Q5: What is the efficiency of co-delivering different AAV particles into the same cells in...any difference in transfection efficiency when delivering these viruses via retro-orbital vs. tail vein...
  8. Targeted Mutagenesis with EvolvR

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    Blog Post
    ...introduced by lambda red recombineering and Multiplex Automated Genome Engineering (MAGE) will always be ...methods for bacterial genome engineering   Learn more about lambda red recombineering of bacteria Use site directed...within a defined region of a genome. Require stringent bacterial growth conditions. Expression of an ...only increasing the standard mutation rate seen during DNA replication by 120-fold over background. Due...
  9. How Addgene Creates Content

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    Blog Post
    ...educational content Addgene truly believes that sharing speeds science, and our entire mission is to enable...informative content to help researchers in creating and sharing knowledge, related to the types of materials we...help.addgene.org, where you can easily search articles answering all of our frequently asked questions!    ...published alongside our new antibody services in the spring of 2022. The first step was to understand information...licensed and cited figures from papers or image sharing repositories. Then we format, do a final review... and knowledge changes and require updating or retiring. CRISPR/Cas9, for example, has gone through rapid...
  10. Viral Vectors 101: Viruses as Biological Tools

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    Blog Post
    ...Varmus H. Retroviruses. Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY, USA: 1997. PubMed...them attractive and tractable tools for genome engineering. Scientists have engineered viruses to target...incompetent viral particles, are required. We will be covering these safety measures in a separate, upcoming ...breakdown of the nuclear envelope, which occurs during mitosis. The most well-known retrovirus is HIV....PubMed PMID: 21433340. 4. Cronin J, et al. “Altering the tropism of lentiviral vectors through pseudotyping...
  11. 3 Challenges in Plant Synthetic Biology

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    Blog Post
    ...and Keira Havens, co-founders of Revolution Bioengineering. Read their previous blog post about how they...directed at GM crops has burdened this emerging engineering technology. Monoculture and chemical spraying... you have developed a genetic tool for plant engineering, please share it with Addgene to both build the...biology community and to support more efficient engineering of a better tomorrow. Thank you to our guest...outside of Washington D.C. and has always enjoyed exploring the outdoors and experiencing nature first-hand...SynBio Reading: Learn more about Revolution Bioengineering Follow RevBio on Twitter Like RevBio on Facebook...
  12. Choosing the B(right)est Fluorescent Protein: Photostability

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    Blog Post
    ...or little deterioration or change of the signal during the course of the experiment. The best fluorescent...photobleaching rates (Shaner et al, 2008). This brings us to the key question: what is the best way to...photostability of different fluorescent proteins? Measuring photostability Since it is clear that a photostable...measurements for different fluorescent proteins and by comparing how the fluorescence intensity changes over time...than predicted by these high power experiments. Measuring photostability at realistic powers will provide... but they do not mimic the natural situation. Measuring photostability in living cells provides a much...
  13. Transferable Skills Guide: Cross-team Communication

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    Blog Post
    ..., too. And remember, if you discover during grad school or during your postdoc that you enjoy and have...engineers were attending the same meetings and hearing the same words, they were leaving these meetings...know what kind of plate they were talking about during the meetings. And “plate” certainly wasn’t the ...experimental protocols. Clarify: Asking for clarification during a meeting can help get everyone on the same page...have come upon any other effective approaches for clearing up communications lines in your professional life...
  14. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

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    Blog Post
    ...Challenges of engineering channelrhodopsins There are three main challenges to engineering channelrhodopsins...channelrhodopsins with high-throughput proteins engineering methods, like directed evolution, because channelrhodopsins...-stimulate” their VTA dopaminergic neurons by triggering a burst of light that could activate ChRger2 ... require fiber-optic cannula implantation, but altering ChRgers so they can be activated with tissue-penetrating...2019) Machine learning-guided channelrhodopsin engineering enables minimally invasive optogenetics. Nat ...
  15. Hot Plasmids and Viral Preps - March 2021

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    ...Here's what you'll find in this post: Epigenetic engineering with single-pot assembly Tracking cell-cell interactions...New items from the viral service Epigenetic engineering with single-pot assembly By Eric Perkins Though...epigenetic researchers custom design their own engineering plasmid tools in about a week. Taking advantage...receiver cells. What’s more, G-baToN can be used to bring cargo, such as protein or DNA, to manipulate function...CRISPR Plasmids and Resources page.  Plasmids for delivering CRISPRi components by a single AAV vector are...Choudhary's lab developed a chemogenetic system that brings Cas9 to ubiquitin ligase for degradation.    ...
  16. Viral Vectors 101: Virus Safety

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    Blog Post
    ...replication of virus and its infectivity in check. Engineering controls, safety systems that are built into ... you can receive appropriate treatment and/or monitoring. Virus-specific concerns   In nature, some viruses...protection! When centrifuging viruses (a necessary step during production), there is a significant risk for aerosol...of following your lab’s biohazard policies. Administering Virus Many virus experiments involve needles...accidents is not following safety protocols.) If administering virus to mice, part of your preparation should...let you work with these valuable tools without endangering yourself or others. Happy transducing! Resources...
  17. Selecting Your Plasmid Purification Kit

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    Blog Post
    ...There are a few different issues that can arise during the plasmid purification process. One is the possibility...broken into a few broad stages: lyse the bacteria harboring your plasmid; separate plasmid DNA from unwanted...environmental impact, check out Addgene’s blog post on bringing sustainable practices for the lab. Plastic consumables...from bottles and the overall weight of the package during shipping. Of the kits reviewed here, NEB’s Monarch..., so be sure to check with your lab manager or ordering department for these.  While budget is an important...Different Uses Plasmids 101: Optimizing Plasmid Yields Bringing Sustainable Practices to the Lab: Easy Action ...resources on addgene.org DNA Quantification Protocol Recovering Plasmid DNA from Bacterial Culture Protocol ...
  18. Common Injection Routes in Mice

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    Blog Post
    ... the abdomen. They are a popular choice for administering different drugs and viral vectors. Like IV injections...suggests, in the muscle. Common uses include administering vaccines, pain meds, or other drugs. Identifying...don’t always require needles. If needles make you cringe or your experiment will allow you to avoid poking...straight to the stomach. This technique involves administering medication through a feeding tube with a bulb...introduction to common injection routes in mice! When considering what method you’d like to use, remember that ...Organisms Additional resources on Addgene.org Genome Engineering Guide CRISPR Mammalian Expression plasmids Cre-Lox...
  19. Light Sheet Fluorescence Microscopy

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    Blog Post
    ...tissue clearing Similar to light microscopy, there have been significant advances in tissue clearing techniques...brightness with increasing depth caused by light scattering and absorption.  Finally, the photomultipliers...techniques during this century.  The first successful attempt to render fixed anatomical preparations transparent...Ultramicroscopy with tetrahydrofuran-based tissue clearing (13,14). A few years later, Karl Deisseroth’s ...  The combination of LSFM with multiple tissue clearing strategies and neuronal labeling methods will ...
  20. 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....
Showing: 321 - 340 of 1016 results