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  1. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

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    Blog Post
    ...know that most cephalopods die after they lay their 1 clutch of eggs? Propagation is thus quite difficult...organism blog posts: Five Popular Model Organisms, Part 1 Five Popular Model Organisms, Part 2 ...promotes scientific sharing The EDGE Program is designed to expand the number of systems that can be studied...genotype to phenotype studies New organismal systems described at this meeting were extraordinarily diverse. ...generating interesting drug-like compounds especially cardenolides which have been used to treat heart disease....disease. Caterpillars that eat milkweed generate cardenolides that tastes yucky (and is poisonous) for bird ...control who a female wants to mate with and if she decides to raise their offspring or leave them to their...
  2. The Addgene Antibody Data Hub

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    Blog Post
    ...an antibody performed in an experiment.   Figure 1: The front page of the Addgene Data Hub.   Antibody...antibody worked under the conditions described above, and a short description of how the antibody performed....the submitted application, along with a brief description of the experiment. If that data was included ...ELISA — but in all cases, it contains a detailed description of the experimental conditions.     Figure 4...The submission process is easy, with simple, descriptive fields, drop-down boxes when appropriate, and...
  3. CRISPR Activation: A Practical Guide

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    Blog Post
    ...activity to a transcriptional activation domain (Fig 1). Two ways to use CRISPRa for your research CRISPRa... Stucka R, Hauber J, Lange UC (2016) Targeted HIV-1 Latency Reversal Using CRISPR/Cas9-Derived Transcriptional...whichever guides are closest to the transcription start site (TSS). We recommend targeting the guides to a ... you should also be ready to test three or four guides directed towards that gene as there can be a large...transcription. That being said, we generally pick guides by putting the promoter of the gene into a gRNA...
  4. Addgene's Website Update

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    Blog Post
    ...straight to your materials of interest!   Figure 1: The new search bar.   Pro tip! You can use a manuscript...and the Addgene Help Center. A big part of this redesign was pulling all of our science options into a ...Addgene’s website easier to navigate, thanks to design choices informed by Web Content Accessibility Guidelines...word for it — check out all the new features and designs addgene.org! And when you’re finished, feel free...
  5. Your Top Requested Plasmid in 2015!

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    Blog Post
    ...forward to another great year of sharing!   References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout...Feng Zhang’s lab at the Broad Institute and was designed and cloned by Neville Sanjana. This fantastic ...your gRNA of interest to mammalian cells. It was designed to have increased lentiviral titer as compared...sampling of the research it has already been used for includes work probing mTOR signaling in cancer cells, deciphering...
  6. Antibodies 101: Reading a Flow Plot

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    Blog Post
    ...Figure 1). Clustered events are called a population.                             Figure 1: While...series of markers are set up sequentially, from broadest to most specific marker, and are represented as...are, try drawing a quadrant over the plot that divides it into four, using the location of the cell populations...Gating and plot orders  As the gates progress from broadest markers to most specific, the first step is to...
  7. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

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    Blog Post
    ...breaks (DSBs) or donor DNA templates.  Figure 1: An overview of prime editing. Created with BioRender.com...demonstrated ~20-50% efficiency in HEK293T cells with 1-10% indels. Since then, further innovations have continued...Engineering. https://doi.org/10.1038/s41551-024-01227-1 Yan, J., Oyler-Castrillo, P., Ravisankar, P., Ward...site by the pegRNA — a guide RNA that also encodes the desired edit and homology to the genomic DNA locus...both specifies the target site and contains the desired edit (or edits) engages the prime editor protein...and the reverse transcriptase domain copies the desired edit, directly polymerizing new DNA onto the nicked...edit within what Fyodor Urnov refers to as “PAM deserts” in the genome. More versatile and precise than...
  8. Inntags: Innovative Protein Epitope Tagging

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    Blog Post
    ...older epitope tags - only time will tell! References 1. Georgieva, Maya V., et al. "Inntags: small self-structured...First described in the 1980s, protein tags are now one of the most useful items in a scientist’s toolbox...IT10 expressed well in cells and did not form undesirable aggregates. Using budding yeast as a model system...
  9. New Viral Vectors - March 2024

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    Blog Post
    ...vectors, and many more, in the table below.    Table 1: New ready-to-use viral vectors in the Addgene repository...pAAV-CAG-DIO-NLS-mRuby3-IRES-eGtACR1-ST AAV9 Optogenetics Adesnik New viral vector pAAV-nEF-Con/Foff DREADD Gi-...
  10. Changing Labor Laws Bring Increased Postdoc Wages

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    Blog Post
    ...$47,476 and the date for implementation was set for Dec 1 2016. In order to be exempted from the need to pay... these institutions (populations potentially from 1 to 5761, according to the NSF data) and therefore ...around $45,000, but as I’ve discussed elsewhere (slides here) there are many postdocs paid at much lower...expected for postdocs in many cases should be as described in the notice of the new NRSA levels and in the...
  11. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

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    Blog Post
    ...making it easier for us to do our work.” References 1. Athanasios Lampropoulos et al. "GreenGate - A Novel...colleagues describe in the PLoS ONE paper that introduced the kit in December, GreenGate includes the six... bring this into your target organism. We have designed a Golden Gate system based on the daily needs ...and cut another,” Lohmann said. “It allows us to design modules that always have the same recognition site...produced will have different overhangs, they can be designed such that they go back together only one way and...
  12. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

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    Blog Post
    ...together along with the Gibson assembly master mix for 1 hour, and then transform into competent cells. The...you should consider giving it a try! References: 1. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison... friends on the Cambridge 2010 iGEM team that describes the basics of Gibson assembly as a parody of “...assembly The Gibson assembly technique was first described by Dr. Daniel Gibson and colleagues at the J. ... Both Snapgene and NEB have tools that help you design primers for PCR amplification of fragments to incorporate...demonstration of how to use Snapgene’s program to design primers for Gibson Assembly. For a simple example...without scars as depicted below. First, you need to design primers to amplify the two fragments while also...
  13. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ...improved sensors for these and other targets. Figure 1: MTRIA biosensors are generated by inserting a circularly-permuted...fluorescent biosensors. Nat Commun. 2022 May 25;13(1):2919. doi: 10.1038/s41467-022-30685-x. Feel the ...Addgene’s catalog is a new class of sensors for neuropeptides and neurotransmitters. Motivated to develop ...GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost A to Z! - from ATP...a new preprint, Jared Toettcher and colleagues describe pYtags, a modular system for monitoring specific... tool for monitoring RTK signaling.  Figure 4: Design of pYtag fluorescent biosensors for monitoring ...iGECInano plasmids here!   Matlashov ME, et al. Design and Initial Characterization of a Small Near-Infrared...
  14. Antibodies 101: Introduction to Gating in Flow Cytometry

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    Blog Post
    ...at a low angle (FSC) or large angle (SSC) (Figure 1). FSC values depend on the cell's size, while SSC ... or ask experienced colleagues.   Figure 1: The laser pulse illuminates the cell and is scattered... your cells based on their size, the cytometer provides you with two measurements: forward scatter (FSC...SSC-H, or FSC-W/SSC-W, respectively. H, A, and W describe the shape of a histogram, which graphically represents... of the cell’s illumination (see Figure 2). H describes the maximum signal strength, while W results from...photocurrent vs. time. The resulting histograms is described through maximum current (H), the time the cell...depends on cell type and laser voltage. The voltage describes an electric potential that can be applied to the...
  15. A Control for All Seasons

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    Blog Post
    ...compare the experimental group.     Figure 1: A, Most biological experiments will benefit from ... Histochemistry & Cytochemistry, vol. 59, no. 1, SAGE Publications, Jan. 2011, pp. 6–12. Crossref,...controls and be able to use this information to design thoughtfully and thoroughly controlled experiments...is generally a sample or group that will have a desired response. For example, if you are studying the ...sample, the antibody binds specifically to its desired target (purple) and nonspecifically through the...The Preimmune serum control will not bind to the desired target but will bind nonspecifically through the...the Fc, allowing you to detect this undesirable interaction. When non-specific interactions are suspected...
  16. Modulate the Activity of 17 Signaling Pathways with One Kit!

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    Blog Post
    ...constructs to the kit in the next two years. References 1. Martz, Colin A., et al. "Systematic identification...so he assembled a plasmid kit designed to do exactly that. As described in the journal Science Signaling...When cancers are treated with drugs designed to hit them right where it hurts, the effects are often ...mutant complementary DNAs (cDNAs) representing core nodes that, when over-expressed, constitutively activated...activators for more cancer-related pathways and more nodes within those pathways. With support from the National...
  17. Your Top Requested Plasmid in 2016!

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    Blog Post
    ...Lab's Nature Protocols Publication describing them here. References 1.Ran, F. Ann, et al. "Genome engineering... and the Puromycin resistance gene as separate peptides. PX459 This is the vector backbone. This particular...Protocols Paper for more information). V2.0 This designation separates pSpCas9(BB)-2A-Puro (PX459) V2.0 from...Guide Pages Browse Our CRISPR Collection Find gRNA Design Tools ...
  18. Fluorescent Proteins 101: When GFP lets you down

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    Blog Post
    ... remains fluorescent in acidic organelles (Figure 1), showing that its acid tolerance is maintained in...follow him on twitter: @joachimgoedhart.   References 1. Zacharias, David A., et al. "Partitioning of lipid-modified...cofactor binding proteins as described above. The smallest tags are peptides that are engineered to bind...homology domains can be used to detect specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production...Péter, and Tamas Balla. "Live cell imaging of phosphoinositides with expressed inositide binding protein domains...
  19. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

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    Blog Post
    ...single neurons in the living brain   Figure 1: Plasmid mix to label neuronal morphology (eYFP) and...genOway press release for more information. References 1. Atasoy, D., Y. Aponte, H. H. Su, and S. M. Sternson... this re-inversion issue. It became possible to design FLEx vectors (Flip-excision vectors) that only ...
  20. CRISPR 101: Off-Target Effects

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    Blog Post
    ...downstream experiments. You only successfully generate 1 null clone and proceed with it. In this case, the ...there are 10 putative off-target locations you can 1) determine if editing at those sites would confound..., it’s essential to understand them in order to design the most specific assays possible. In this blog...-target effects should impact your experimental design differently. Yes, it is always better to reduce...introducing a specified edit). However, if your design does have wiggle room, you can minimize off targets...
  21. Microbes: Look, Listen, and Tell

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    Blog Post
    ...communication check out these SciComm blog posts. References 1. Bourdeau, Raymond W., et al. "Acoustic reporter genes...is absolutely fascinating to see the living art designed by creative minds with a (I assume) very steady...Our Microbiology Resource Page CRISPR plasmids designed for use in bacteria Deposit Your Microbiology ...
  22. Fluorescent Proteins 101: History of Fluorescent Proteins

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    Blog Post
    ...in the crystal jellyfish Aequorea victoria (Figure 1) in molecular detail. On the underside of the “umbrella...characteristic radiance for some time – there are records describing them dating back to the first century A.D. – ...mechanistic understanding of GFP’s function. His lab described the protein’s structure and also discovered many...
  23. SciComm with the Experts at Science in the News Part 2

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    Blog Post
    ...to the interview with Science in the News Chapter 1: Introduction - Vini and Amy tell us a little about... discusses some of the programs SITN runs and describes some of the more popular topics from the Science... fields as well as students from the ed school, design school, and Kennedy school to discuss the impacts...impacts of science in a social context. Episodes are driven by narrative and story telling. Chapter 5: The...The SITN Blog - Vini and Amy describe how the SITN blog is a source of scientific information for the ...underwater? Chapter 8: Work-Life Balance - Vini describes how she balances working in the lab and working...
  24. Why Do I Need an MTA Anyway?

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    Blog Post
    ...is here to help. Contact us at [email protected] or +1 617-225-9000, or check out our MTA reference pages...University Technology Managers (AUTM). This MTA provides many benefits for both the providing and receiving...
  25. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

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    Blog Post
    ...normal size.      References: 1. Doench, John G., et al. "Optimized sgRNA design to maximize activity and ... producing a wall of results that can only be described as bigly. But such a project should not be undertaken...Library Screening Why the kinome? While properly designed perturbations – in the case of screens using CRISPR...with CRISPR/Cas9 Get John Doench's Tips on gRNA Design Check Out News Ways to Use CRISPR for Single Base...
  26. Three Tips to Organize Your Lab Notebooks in the New Year

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    Blog Post
    ...organized and boost your productivity in 2014. Tip 1: Use color coding, labels, and post-its to make finding...lab earlier, to read that stack of papers on your desk, and to tackle some new experiments before the next...gathering that pile of lab notebooks and binder on your desk. Arrange them in chronological order (or by project...labels. Color code your Post-its and write brief descriptions on each. Place the notes sticking up out of ...colleagues. Start small by identifying the protocols, guides, or references that your labmates ask to borrow...
  27. Promoters

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    Guide
    ...Strong mammalian promoter from human elongation factor 1 alpha CAG Constitutive Strong hybrid mammalian promoter...gene. The coding strand is the DNA strand that encodes codons and whose sequence corresponds to the mRNA...proximal promoter, and distal promoter. Below describes the specifics of these regions in eukaryotic cells...Pribnow box which usually consists of the six nucleotides, TATAAT. Proximal Promoter Further upstream from...Promoters Used in Research Promoter Expression Description CMV Constitutive Strong mammalian promoter from...Promoters used in Research Promoter Expression Description T7 Constitutive but requires T7 RNA polymerase...
  28. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ...base editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated with...or one pyrimidine base to the other. Figure 1: Base transition editors catalyze purine-to-purine...engineering and protein delivery. Nature Communications, 8(1), 15790. https://doi.org/10.1038/ncomms15790 Richter...DNA bases without breaking the DNA backbone, sidestepping the problem of HDR vs. NHEJ entirely. The first...fourth-generation base editors, BE4, reduce the undesired C→G or C→A conversions that can happen with BE3s...named ABEmax. In 2020, two papers were published describing additional ABEs evolved from ABE7.10 that have...cytosine and adenine base editing, called CABE-Ts and TadDEs without additional fused components (Lam et al....
  29. Easing the Protein Purification Process with pCri

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    Blog Post
    ...individually or as a complete kit from Addgene. References 1. Goulas, Theodoros, et al. "The pCri system: a vector...subtilis and Pichia pastoris pCri also includes plasmids designed for protein expression in systems other...construct, most often in special E. coli strains designed for protein expression. Cultures are then lysed...plasmids are divided into two sets, each of which includes plasmids with different varieties of tags, both...other eukaryotes better than E. coli. Plasmids designed for these systems may include a signal peptide...can improve disulfide bond formation. pCri also includes a special feature for difficult-to-purify membrane...membrane proteins. pCri vector pCri-13a, designed for use in B. subtilis, contains the MISTIC (membrane-...
  30. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

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    ...target cell line or organism (Presented in figure 1). More information on Agrobacterium-mediated transformation...mediated cloning of your gRNA of interest. Figure 1: Simplified schematic of agrobacterium-mediated transformation... While it is certainly possible to select for desirable traits using traditional plant breeding approaches...modification of known or suspected genes that regulate a desired phenotype. In fact, CRISPR has already been used...wheat, mushroom, and rice (Khatodia et al., 2016). Despite the almost universal functionality of the CRISPR...genes in plants using the same general experimental design principles developed in other model organisms (...nanostructures (Zhang et al., 2019) or cell-penetrating peptides (Santana et al., 2020) and plants viruses (barley...
  31. Hot Plasmids: Spring 2025

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    ...synapses and deep-brain fiber photometry.  Figure 1: Characterization of iGluSnFR4 in mice. A) Scheme ...CycT1 or PolII-CTD antibodies and unlabeled MS in DLD-1 cells (colorectal cancer). Image courtesy of Brianna...Glutamate-Sensing Fluorescent Reporter (iGluSnFR) designed and improved by the Loren Looger and Kaspar Podgorski...Aggarwal et al., 2025).In this latest work, the team designed and screened a large library of iGluSnFR3 mutants... HA-, and EGFP-expressing lentiviral backbones designed right here at Addgene. Here's what makes these... stable cell line creation a breeze. Modular by design: Key elements (promoter, tag, polyA site) are flanked... plasmids for flexibility and speed, so you can design, test, and innovate faster than ever. Plus, we'...
  32. Overcoming the AAV Size Limitation for CRISPR Delivery

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    ...science.aaq0180  Fine EJ, Appleton CM, White DE, Brown MT, Deshmukh H, Kemp ML, Bao G (2015) Trans-spliced ...indel formation and phenotypic changes as early as 1 week post-injection. Livers from these mice were histologically...as potent and specific? The two AAV strategies described above showed successful target modification, indicating...from Streptococcus thermophilus and a rationally-designed truncated Cas9. Unfortunately, certain drawbacks... approximately 1350 amino acids in size, which includes SpCas9, and shortern orthologs approximately 1000...Split inteins are a pair of naturally occuring polypeptides that, when at the ends of two proteins, mediates... proof-of-concept split intein SpCas9 which had modest editing rates in HEK-293T cells when compared to...
  33. Bringing Sustainable Practices to the Lab: Innovations

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    Blog Post
    ...products by their environmental impact on a scale of 1-10: the lower the score, the lower the environmental...collect in those red biohazard bags, typically is destined for a landfill. However, Triumvirate Environmental...Then, they use this material to produce various grades of plastic lumber and distribute it to industrial...
  34. Our Most Popular Blog Posts from 2019

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    ...deletions, and all possible base-to-base changes.   1. Plasmids 101: Transformation, Transduction, Bacterial...on the Addgene blog and have created and updated guides on various topics in biology over the past year...containing these fluorescent proteins at Addgene. 7. Designing Your Chalk Talk for the Academic Job Interview...
  35. 6 Steps to Submitting a Resume That Gets Seen

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    ...resume that will give you a chance at an interview:  1) Have an excellent resume ready to be submitted The...experience. Most Addgene PhD level job descriptions clearly describe that the roles do not involve bench ...submission of a job application. 3) Read job descriptions carefully Be open minded and be bold. You don...qualifications to apply for a position.  A job description is usually a wish list and the hiring manager... way to a hiring manager. Learn to confidently describe your strengths in your resume and don't be shy...
  36. Antibodies 101: The Basics of Western Blotting

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    ...to western blotting  In a western, proteins are: (1) separated by size, (2) transferred to a membrane,...  (3) detected using antibodies.   Figure 1: Overview of the Western blot process. The sample,...protein analyses using ELISA. Peptides 72:4–15 . https://doi.org/10.1016/j.peptides.2015.04.012 Burnette WN ... Figure 2: The western blot dry apparatus includes a stack of filter paper, a copper anode/iblot ...blocking buffers, and incubation times. If the vendor includes publications that cited the antibody, consider...abundance In addition, the thickness of the band provides information about the relative abundance of the...
  37. Addgene's Top Blog Posts from 2020

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    Blog Post
    ...five methods for extracting DNA without a kit too. 1. Early Career Researcher Toolbox: Free Online Molecular...databases, DNA sequence manipulation, and primer design. Best wishes for the new year and your research...
  38. Top 10 Open Science Developments of 2013

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    Blog Post
    ...2013 edition. 10 new developments in open science 1)      In February, the Office of Science and Technology...     Science’s John Bohannon published an article describing a sting operation in which he submitted a phony...
  39. Technologies Enabled by NanoLuc® Luciferase

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    Blog Post
    ...furimazine by Promega’s Chemistry group is a good story (1). From a deep sea shrimp to a compact powerhouse of...cellular analysis products at Promega.     References 1. Hall, M.P., et al. (2012) Engineered luciferase reporter...above 600 nm. LumiFluors Schaub, F.X., et al. (3) designed two imaging probes. One consists of NLuc fused...Reporters. The LumiFluor reporter constructs were designed to be packaged into a retrovirus for infection...BRET probe for imaging. These authors set out to design a better fluorescent protein that could be co-imaged...acceptor, the more powerful the FRET signal. In well-designed FRET-based biosensors, the intensity of the FRET...
  40. Finding Your Science Policy Path

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    Blog Post
    ...relations, advocacy, and writing legislation (see Figure 1 for career map). The beautiful part of science policy...about science policy as a profession. Figure 1: Science policy can lead you to many paths! For a ...uncertainty of it all. Though I had spent the first few decades of my life following a very straightforward conveyor... harassment in academia or a large company who decides how they are going pursue regulatory approval for...their Local Engagement Playbook for step-by-step guides on how to engage in your local community.  Read...comprehensive resource developed by Dr. Kelly Singel which provides an overview of all available science policy fellowships...
  41. Tag Your Favorite Yeast Genes with Ease

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    Blog Post
    ..., or CaURA3 selection marker. Lee et al.(1) assessed many of these fluorescent tags in S. cerevisiae, ...protein products is desired, as multiple selection markers can ensure that all desired tags have been integrated...integrated. Simply design your amplification primers with the desired targeting homology—in frame, of ...These tagging vectors are based on previously described pFA6a-link vectors and include a Kan, SpHIS5, ...former lab at UNC Chapel Hill. Bähler et al.(2) describe a modular collection of plasmids for a wide variety...3xHA, 13xMyc, GST, or GFP). Longtine et al.(3) describe a complimentary set of plasmids for use in S. ...
  42. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

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    Blog Post
    ...depends on what you are looking to study. Figure 1: A schematic showing the differences in chemogenetics... Roth BL (2015) DREADDs (Designer Receptors Exclusively Activated by Designer Drugs): Chemogenetic Tools...less invasive and more flexible as it relies on designer ligands that can be simply introduced by injection...lab and Takafumi Minamimoto’s lab known as deschloroclozapine (DCZ) has been developed to meet these requirements...should be carefully controlled for in experimental design.   Optogenetics Chemogenetics Timing Fast ...Find our chemogenetics and optogenetics science guides ...
  43. Phage Directory: From Phage Therapy to a Repository of Phage Information

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    Blog Post
    ...information Since the inception of Phage Directory ~1 year ago, Sacher and Zheng found that they could expand...infection, she shared that tweet with Jan Zheng, a UX designer that she knew. “We had actually met at a lindy...to possess the correct phages will test. These includes labs with a large number of different phages or...
  44. Six Spooky Science Stories and Halloween at Addgene

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    Blog Post
    ...Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID: 15026169. Soo, Rochelle M., et al...wall of green algae (Soo et al., 2015). There, it divides into two cells. At the same time, it sucks out ...1983). Grasshopper parasite The parasitic worm Spinochordodes tellinii‘s larvae makes its home inside grasshoppers...
  45. New CRISPR Web Resources and #12DaysofCRISPR Recap

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    Blog Post
    ...problem - see our recap below! Day 1: gRNA design tools: How to Design your gRNA for CRISPR Genome Editing...given our CRISPR Plasmids and Resources a fresh new design, sorting the plasmids we have available by category...capabilities. Epigenetics: CRISPR has made it possible to design programmable epigenome editing tools, and the field...cleavage in bacteria. In mammalian systems, Cas13a degrades only the specified RNA target. Bonus: It displays...
  46. Build Your CRISPR Vocabulary

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    Blog Post
    ...  Figure 1: Overview of the parts of CRISPR. The bacterial chromosome encodes a tracrRNA (in some...DNA sequences. In laboratory settings, an sgRNA includes the crRNA and tracrRNA sequences in a “single-... sequences in the CRISPR array that will become guides for future targeted degradation. Before integration...biology of Cas9. Want a simpler guide to keep at your desk? We’ve got a CRISPR cheat sheet for that! Resources...
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