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  1. Synthetic Biology, Artificial Chromosomes, iGEM & More - Interview with Tom Ellis

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    Blog Post
    ... grad school to a startup and back to academia. Chapter 1: How working at a startup prepared Tom for an...synthetic biology, synthetic chromosomes, and obstacles to becoming an academic researcher. Listen to...to the UK with foundational research, training, startups, and real applications of SynBio. Chapter 3: Synthetic...of biology at the level of DNA using modular, standardized, disciplined, and open techniques. Chapter 4...Golden Gate Cloning Learn About Working at a SynBio Startup in Our Interview with Michael Koeris Resources...
  2. 500,000 Plasmids Shipped and Counting!

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    Blog Post
    ...as transformed bacteria in stab culture format. Our lab team creates stabs for each plasmid being shipped...information in our lab software, labeling stabs with barcodes as they go. Stabs are organized by shipment and moved...their boxes with shipping labels and paperwork. Our stacks of Addgene boxes are now ready to be sent around...depicts where plasmids have been shipped to. Plasmids start out at our Cambridge office – previously at 1 Kendall...
  3. How Exercise Purges Stress and Keeps Depression at Bay

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    Blog Post
    ...only in their skeletal muscle show resistance to stressful circumstances – including unpredictable loud noises...mck-PGC-1a1 mice, and those who exercise, show resistance to depression. For the researchers led by Dr...hypothesis was that trained muscle would produce a substance with beneficial effects on the brain,” Ruas said...produces an enzyme that purges the body of harmful substances. So in this context the muscle's function is ...your own lab, you’ve come to the right place. Get started with PGC-1 plasmids from Addgene. References: ...
  4. Your Top Requested Plasmid in 2016!

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    Blog Post
    ...fused to the puromycin resistance gene through a 2A peptide. The puromycin resistance gene enables selection...uninitiated, contains much of the information needed to understand how to use it. TLDR: This is a versatile mammalian...ribosome to translate Cas9-2A and the Puromycin resistance gene as separate peptides. PX459 This is the ..., which had a point mutation in the puromycin resistance gene that made it less effective in certain cell...nickase), which nicks DNA rather than creating double stand breaks and may better facilitate homology directed...
  5. What's New in CRISPR - Spring 2019

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    Blog Post
    ...String assembly gRNA cloning (STAgR) String assembly gRNA cloning (STAgR), developed by Stefan Stricker...developed another AAV toolkit - this time using the Staphylococcus aureus Cas9 (SaCas9). This AAV-based genome...high numbers of gRNAs in one overnight reaction. STAgR is highly customizable as vector backbones, and ...chosen. Read the paper in PLoS One Find the STAgR cloning plasmids Mobile-CRISPRi allows CRISPRi...from various environments. It has the advantage of stable genomic integration and is easily transferred via...
  6. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ...voltage response, brightness, and photostability. It enables sustained deep-tissue imaging in vivo. Image... and increased brightness, sensitivity, and photostability under two-photon illumination than previous...pAAV-EF1a-DIO-JEDI-2P-Kv-WPRE   Liu Z, et al. Sustained deep-tissue voltage recording using a fast indicator...ZAP70 (ZtSH2) which binds only the phosphorylated state of an immunoreceptor tyrosinase-based activation...options for potassium biosensors and are a promising starting point for further engineering. Figure 5: Structure-guided...lifetime imaging. B) LiLac responses are highly resistant to pH changes. Image adapted from Koveal et al...
  7. Site Directed Mutagenesis by PCR

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    Blog Post
    ...facilitate the repair, the template should be resistant to Cas9 cleavage. If a plasmid contains the template...cleavage), thereby rendering the resulting construct resistant to Cas9 induced cleavage. Summary of Site Directed...and bacteria are transformed with the nuclease-resistant nicked plasmid (the PCR product). Plasmids are... pieces of double stranded, circular DNA with staggered nicks, recent data (see Xia et al., 2015) suggests...products into will efficiently patch up the DNA. The resistance marker from the parental plasmid provides a mean...overhangs and ligate these blunt ends together in a standard ligation reaction resulting in a frame-shifted...Breslauer, Kenneth J., et al. "Predicting DNA duplex stability from the base sequence." Proceedings of the National...
  8. Science as Art

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    Blog Post
    ...based at the University of Queensland, whose outstanding marine photography has attracted much attention...what science is about. For me it is essential to stay in touch with the living organism I study, to be...So photography can be used as a way to better understand and appreciate the organism, for both scientist...Geographic and Smithsonian Magazine, where the editor stated “Albert G. Richards, a teacher of dental radiography...on teeth, but on blossoms, thus revealing the startling inner beauty of familiar flowers." This new x-...departments to showcase the work of students and staff by holding scientific art competitions where creations...
  9. Three Tips to Organize Your Lab Notebooks in the New Year

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    Blog Post
    ...frustrated and no closer to starting your experiments. Get organized now! Start by gathering that pile of...wake up and get to the lab earlier, to read that stack of papers on your desk, and to tackle some new experiments...or projects) to make it easier to know where to start your search. Then identify the most frequently used... and resource investment required to change the status quo. But with recent improvements to ELN and online...ways to share your documents with your colleagues. Start small by identifying the protocols, guides, or references...
  10. IBEX Knowledge-Base: A Data Resource for Multiplex Tissue Imaging

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    Blog Post
    ...conventional microscopes, amplified staining methods, and oligonucleotide-based staining. Of course, if you’re interested...multiplexed tissue imaging method that uses iterative staining and chemical bleaching to allow for high-resolution...reagents that you know work…preferably before you start your experiment. (And you’re certainly not alone...Biomolecular Atlas Program (HuBMAP) antibody metadata standards. And the dataset is rich. Experimental data...seeking to create an iterative process that is constantly becoming better, with more data, more discussion...
  11. What's New in CRISPR - September 2019

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    ... from an analysis of human metastasis cohorts and a prior mouse metastasis CRISPR screen. The Chen Mouse... Using the structure of CasX at eight different states of assembly from cryo-electron microscopy, the ...mutated human estrogen receptor. This protein fusion stays in the cytoplasm, but with the addition of 4-hydroxytamoxifen...knockout libraries for cancer research. The Mouse Metastasis CRISPR-Cpf1(Cas12a) Double Knockout Library is...tumor suppressor genes (TSG). Find the Mouse Metastasis CRISPR-Cpf1 Double Knockout Library Find the...
  12. pSiM24: Simplifying Plant Genetic Engineering

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    Blog Post
    ...continue to improve plant nutrition, yield, and pest resistance. Much of plant genetic engineering revolves around...optimize this system for gene transfer, studying the stability of modified Ti plasmids during plant infection...versatile binary Ti vector for both transient and stable gene expression applications in plants. This smaller... To build a better binary vector, Sahoo et al. started with a vector commonly used by their lab, the 13...variety of plant species. The kanamycin antibiotic resistance gene is controlled by a weaker constitutive Nos...expression vector for transient assays as well as stable expression of foreign genes in plants." PLoS one...
  13. Bricking Science: Portraying Scientific Reality Through LEGO

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    Blog Post
    ...process One major challenge when we started was creating a LEGO stage that resembles a real laboratory. ... pathways just waiting to be discovered. As we started building, we thought – how can we make the connection...in a custom made, colorful laboratory. When we started, the hardest part of the process was to embed personality... trained to follow protocols, guidelines, and standards. Bricking science reminded us of our inner children...children who have no preconceived obstacles, optimistically grasp at limitless potential, and who sometimes...
  14. Fostering communication in remote collaborations

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    Blog Post
    ... of the project. The state of research is constantly shifting, and long-distance collaborations are becoming...between each meeting, but short enough that everyone stays on the same page. Ideally, these meetings include...Conference in Frankfurt, Germany When funding and circumstances permit, think about having collaborators spend...communication within an existing collaboration, or start something new! Additional resources on the Addgene...
  15. Plasmids 101: What is a plasmid?

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    Blog Post
    ...new to plasmids may need some extra guidance to understand the specific components that make up a plasmid...and techniques, and empower you with a firm understanding of the fundamentals. Our mission is to curate... more functional benefits to the host such as resistance to antibiotics, degradative functions, and/or...advantageous for survival, such as an antibiotic resistance gene. In contrast, plasmids utilized in the ...recruiting replication machinery proteins Antibiotic Resistance Gene Allows for selection of plasmid-containing... obtained. Selectable Marker The antibiotic resistance gene allows for selection in bacteria. However...
  16. Viral Vectors 101: Parts of the AAV Packaging Plasmid

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    ...AAV.  Where do you start? Plasmids of course! You just need three plasmids to start making the AAVs you...endonuclease domain. Following replication, Rep78/68 stays attached to the AAV genome and docks with the genome...Alternative splicing and differing translational start sites generate the three VP proteins. The three ...clearly defined yet. Now you’ve got a basic understanding of the AAV RepCap plasmid! Check out this blog.../doi.org/10.1007/s40259-017-0234-5  Büning H, Srivastava A (2019) Capsid Modifications for Targeting and...
  17. The Stingy Scientist: How the Baby Gel Box Was Born

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    ...was using.  I started to do little things to save reagents and money.  This was the start of my interest...resources where possible.  At the lunch table, for instance, we have a plastic container problem…reusable ... a popular topic in the media.  At one point I started to become very aware of how little care I took ...time and money. My other stingy science project started when I realized I was pouring large agarose gels... a small portion of the gel.  What a waste?  I started looking for smaller and smaller gel apparatus until...
  18. Viral Vectors 101: Optogenetic Tools

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    Blog Post
    ...turn on with a light-pulse at one wavelength and stay on until a second pulse of light, with a different...it off, allowing the cell to be excited over a sustained period of time (Gong et al., 2020). Light-gated...signaling cascade that may go awry in a disease state.      Figure 4: Example of an intracellular...optogenetics or biosensors in one system. There can be substantial overlap in the activation curves (Marshel et ...system in your own work, you’ll need to know (and understand!) the hardware types, construction, and testing...main text.  Then, check out the Deisseroth lab’s Stanford Optogenetics Resources and the Boyden lab’s Synthetic...havoc on your experiments, so it is important to understand the details of how each tool performs and the...
  19. Viral Vectors 101: AAV Variables That Matter

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    ...just dive right in and start injecting your virus, a successful AAV experiment starts with validation and...your system that can affect expression, before starting the selection process. Factors to think about ...expression.   Once you know that, you’re ready to start selecting, testing, and optimizing three key variables...serotype that works great in a GABA neuron, for instance, may not transduce the adjacent norepinephrine...in your system and then test each vector before starting your experiments (Rolland et al., 2016; Su et ...selection …is truly validation and optimization! Understanding serotypes, promoters, and titers will allow ...
  20. 10 Great Guest Posts We're Thankful For

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    Blog Post
    ...Addgene Blog? Fill out our guest blogger form to get started!   Thanks for all your hard work!   Fluorophores... Nucleic acid based machines Recombinase-based State Machines Enable Order-dependent Logic in vivo Nathaniel...received by a cell. These so-called recombinase-based state machines could potentially be used to report upon...this post to learn more about recombinase-based state machines and how they function. Adapting Toehold...important things to consider if you’re getting started with them. CRISPR Pairing CombiGEM and CRISPR ...efficient genome editing - all influenced by an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking...
Showing: 441 - 460 of 1028 results