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Showing: 421 - 440 of 653 results
  1. From our Table to Yours: An Inside Look at Lunch at Addgene

    Type
    Blog Post
    ...coveted. I’ve only won 2 trophies over the years, which is either an indication of the stiff competition...building activity that is easy to organize. We’ll dedicate a post to these competitions in the future, and...establish, with the goal of fostering better communication between labs. Scientists are often willing to...
  2. Six Spooky Science Stories and Halloween at Addgene

    Type
    Blog Post
    ...melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID: 15026169. Soo, Rochelle... into a pulsating display imitating a moving caterpillar. The worm controls the snail’s behavior, forcing...exposed to predators. Hungry birds spots the “caterpillar” and then poke out the snail’s eyes and ingests...
  3. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    ...elution choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose ...If you use a kit for DNA purification or if you use a DIY purification protocol, you might have noticed... mutations in single stranded DNA undergoing replication.   DNase on the other hand can digest and degrade...degradation during plasmid DNA extraction and purification from E. coli is the presence of the protein ...most common E. coli strains used for plasmid amplification are endA negative, meaning the gene has been...chelating agents that can be introduced in the purification protocol to remove endonucleases activity. For...controls the pH, while the EDTA chelates any divalent cations like Mg2+, preventing the activity of DNase. While...
  4. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    ...et al.(2) describe a modular collection of plasmids for a wide variety of genome modifications in S. Pombe...insert will typically contain both a protein modification sequence and a selectable gene product for isolation...tags have been integrated. Simply design your amplification primers with the desired targeting homology—.... cerevisiae, looking at their performance in categories such as brightness, stability, and disruption...optimal FP combinations for use in yeast imaging, categorized by specific filter sets and experimental output...This collection is ideal for tandem affinity purification of protein complexes. John Pringle and Jürg ...deposited a large collection of plasmids for genome modification in yeast developed by Dr. Pringle’s former lab...
  5. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    ...Agrobacterium infection. At approximately 7 kb, pSiM24 is 2 and 6 kb smaller than commercially available pCAMBIA...for both transient and stable gene expression applications in plants. This smaller, easily customizable...Agrobacterium are termed binary vectors because they must replicate in two species: Agrobacterium and E. coli. Like.... coli. However, they also need to be able to replicate in Agrobacterium to ensure that they are maintained... Thus, binary vectors contain two origins of replication, one each for E. coli and Agrobacterium, along...bases each. By switching the E. coli origin of replication and codon-optimizing the Agrobacterium ORI, they...constitutively active M24 promoter has been modified with duplicated enhancer domains and characterized in studies...
  6. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

    Type
    Blog Post
    ...might otherwise be hard to identify. Figure 2: Users can cross-interrogate screens from the literature...   Figure 3: Each publication featured in BioGRID ORCS has a dedicated summary page with PubMed...favorite gene was described in a CRISPR screen publication. You see this mentioned in the results section...manually curated and annotated from more than 45 publications, beginning with the seminal research articles...screen curated in ORCS is linked to the original publication via its PubMed identifier, which can also be ...a list of individual screens curated for the publication. Clicking on the screen name takes users to a... supplemental data tables available with the publication are also provided for the user. This approach...
  7. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    ...Nat Methods. 2009 Feb 6(2):131-3.  PubMed. Find plasmids from this publication at Addgene. Subach OM...Since the first research applications of GFP were published in the 1990s, biologists have spent a lot...color was observed after excitation at 488 nm, indicating that spontaneous conversion did not occur. Tobacco... Sattarzadeh et al., Sci Rep. 2015 CC BY 4.0 Applications and limitations of GFP photoconversion One concern...nmeth.1664. PubMed. Find plasmids from this publication at Addgene.   Addgene’s Fluorescent Protein...
  8. New CRISPR Web Resources and #12DaysofCRISPR Recap

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    Blog Post
    ...Genome Editing and The CRISPR Software Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed...job - we strive to also give you high-quality educational resources to help you do better research. CRISPR...design, sorting the plasmids we have available by category. Each plasmid page now has a helpful graphic to... newest page in the Transcriptional Regulation category already has five blue flame plasmids covering ...providing an avenue for temporary/reversible CRISPR modification. In addition to these subpages, our updated ... HypaCas9 Day 10: Transcriptional regulation: Truncated gRNAs for Regulating Gene Expression and CRISPR...
  9. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    ...E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference between arg1 and arg2 vesicles...Knowing where bacteria are located within their host is often key to understanding their role in both...reporters are not as well suited for imaging organs located deep in the body, like the gastrointestinal tract...lab recently discovered that gas vesicles also scatter sound waves and therefore produce signals in ultrasound... experiment to demonstrate the most exciting application for ARG’s: in vivo imaging. Specifically, arg1...
  10. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...posts: Part 1: Genetically Engineered Mice and Part 2: Breeding and Crossing Mice. My first CRISPR mouse...is less predictable. In our best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA...interested in using CRISPR to develop therapeutic applications that better human health.     References Cong...
  11. What's New in CRISPR - March 2020

    Type
    Blog Post
    ...Enhanced CRISPR-based DNA demethylation Figure 2: The components of Casilio-ME1. Image from Taghbalout...expression of specific Caulobacter genes using a catalytically dead Cas9 (dCas9) derived from Streptococcus...methylation-silenced genes. Read the paper in Nature Communications Find Casilio-ME plasmids If you have a new...
  12. Cre-ating New Methods for Site-specific Recombination in Drosophila

    Type
    Blog Post
    ...CHO) cells, as well as in cortical neurons in layer 2/3 of the mouse brain. Rubin’s work shows that the ...Gerald Rubin’s lab sought to make complex genome modifications in Drosophila using multiple recombinases. To...offspring carried both the recombinase and GAL4, indicating that the recombinases were inherited normally...
  13. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...Joung 2016). Table 2: Unbiased Genotyping Options Technique Detection Limit Application Advantages Disadvantages...PMID: 23792628. PubMed Central PMCID: PMC3773023. 2. Vouillot, Léna, Aurore Thélie, and Nicolas Pollet...off-target sites across the genome and pinpoint the location of mismatches based on the sequences of the genome... Figure 1: Schematic representations of the indicated unbiased genotyping options. Figure reproduced...those relating to DNA structure (e.g. epigenetic modification, bulges) that may also impact off-target edits...cell-based methods are genome-wide, unbiased identification of double-strand breaks (DSBs) evaluated by... HTGTS Not reported Cell-based Identifies translocations Limited by chromatin configuration, produces...
  14. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...six different research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (5), Wang/Lander/Sabatini...Bioinformatics 33.7 (2016): 1099-1100. PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to...the user. This approach is useful for several applications as will be discussed below. The software could...contain multiple gRNAs targeting all genes in the indicated species and were suggested to be the best gRNAs...
  15. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    ...their required qualifications and skill sets Taking the time to identify and research 2 – 3 potential...Effective verbal and written communication skills are key to publication and acquisition of funding. If...  Writing Communication Publicity Consulting internships Communication Teamwork Management...Models Communication Regulatory knowledge Networking Blogging Writing Communication Networking...scientists identify potential career pathways, education and mastering new skill sets must occur by doing... skills such as leadership, management, and communication. A great approach to fully understand and acquire... science class: Science Club for Girls) Communication to diverse audiences Organization Mentorship...
  16. Early Career Researcher Toolbox: Free Tools for Making Scientific Graphics

    Type
    Blog Post
    ...project my team & I have poured our hearts into for 2 yrs) has 200K+ figures by 95K+ scientists 👨🏽‍🔬👩‍...software that let me create graphics that better communicated the science in my blog posts. This post highlights...free tools which will hopefully also help you communicate your science, whether it’s in presentations, ...number of drawing tools, so it’s harder to draw intricate figures, like a brain or a mouse. Vectr Vectr ...without the watermark, which are better suited for publications, if you sign up for a paid account.  I’m proud... free medical images organized into four main categories: anatomy and the human body, cellular biology...appropriate credit, provide a link to the license, and indicate if changes were made to the images. This requirement...
  17. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    ... Figure 2: Dense (c), intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, ... use the GAL4-UAS system along with precisely truncated attP sites to control recombination of the gene...of PhiC31. In the SPARC system, attP sites are located on either side of a stop cassette, upstream of ...amount of labeling, you can alter the attP site. Truncating the attP site upstream of the stop cassette results...population is directly correlated to the length of the truncated attP site. The three SPARC variants, based on ...lobe. Image from Isaacman-Beck et al., 2019. Applications for SPARC SPARC technology is versatile and ...dendrites of T5 neurons by placing the calcium indicator GCaMP6f in the effector position, as well as to...
  18. Plan Your Experiment

    Type
    Guide
    ...either (1) a lack of gRNA and/or Cas9 expression or (2) a lack of efficient target cleavage in cells expressing...Manipulation Select Expression System Design gRNA Publications Get Started CRISPR is a powerful system that...may not be a perfect plasmid for your specific application, and in such cases, it may be necessary to customize...reagent to suit your needs. Genetic Manipulation Application Cas9 gRNA Additional Considerations Knockout ...method Expression System Components of System Application Mammalian expression vector Cas enzyme promoter...gRNA complex, which is delivered to cells using cationic lipids Transient expression of CRISPR components...gene you target will depend on your specific application. For example: To activate or repress a target...
  19. 10 Ways to Share Your Science!

    Type
    Blog Post
    ...but also something about how science really works. 2)   Read science writing in magazines or newspapers... Foundation Round Table. In any field, good communication skills help when it comes to finding a job. ...an argument for taking the time to hone your communication skills. So, where can a graduate student or ...opportunities than ever before for scientists to communicate about their work and their interest in science...your area or at your institution; the news or communications office is a good place to start. It’s not unusual...about science writing as a career, the Science Communication Program at the University of California, Santa... your research stories. Consider practicing communicating your research by guest blogging here, just leave...
Showing: 421 - 440 of 653 results