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  1. Plasmids 101: 5 factors to help you choose the right cloning method

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    Blog Post
    ...consider Gibson cloning. In this technique, you first perform a round of PCR to generate DNA fragments with 15...your insert, DpnI digest away the templates, and transform the PCR products into E. coli. Ligases within ...available cloning kits that one could choose from to perform these experiments. In most cases, there will be...
  2. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

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    Blog Post
    ...thousands of designs in parallel. To do this, we performed high-throughput ‘self-targeting’ screens with ...PRIDICT2.0 score. For the best prime editing performance, we suggest that users select some of the best...learning–based model with high generalization performance. Science Advances, 5(11), eaax9249. https://doi.org...
  3. Experimenting with New Careers while in Grad School

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    Blog Post
    ...submitted was hilarious, full of interesting information, and received a lot of attention on social media...Museum and others when I was trying to find local informal science educators, and ever since, the State Museum...the network I built during graduate school is my informal advisory board. Although my goals may not have...
  4. Genome-wide Screening Using CRISPR

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    Blog Post
    ...discuss how CRISPR libraries are being used to perform genome-wide screens and highlight some of the reagents...What are the steps involved in a CRISPR screen? Performing a forward genetic screen using CRISPR libraries...your phenotype of interest. For more detailed information on CRISPR screens check out Addgene’s 2016 publication...
  5. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

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    Blog Post
    ... up after trying out the new protocols.io beta platform for sharing, annotating, and storing life science...associated protocols. When you upload your plasmid information, you’ll have the option to add additional documents...restriction digest of plasmid DNA, and bacterial transformation. And more plasmid-related educational resources...
  6. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...resulted in 15% C-to-U editing activity. They then performed sixteen rounds of directed evolution across the... cytidine deaminase activity. While RESCUE can perform C-to-U edits, the authors note that it retains ...whole-transcriptome RNA-sequencing, the authors performed rational mutagenesis and identified that a S375A...
  7. Plasmids 101: Shuttle Vectors

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    Blog Post
    ...bacterial ones. The mammalian analogue of bacterial transformation is transfection. In stable transfection, DNA...focused this post on shuttle vectors delivered by transformation and transfection and left the business of transduction...Resources on the Addgene blog Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection...
  8. Lab to Office Culture Shock

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    Blog Post
    ...of time that doesn’t involve critical thinking. Former Addgenie Jessica Welch says, “[It] seems counter-intuitive... yourself a Scientist. You work in a lab. You formulate hypotheses and conduct experiments. You transfer...work environments outside of academia provide a formal training regime and the right set of tools to help...
  9. To Codon Optimize or Not: That is the Question

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    Blog Post
    ...protein. Codon optimization could affect protein conformation, folding and stability, change post-translational...  In addition codon optimization could remove information encoded in the primary sequence of a particular...updated resource for codon usage tables. BMC Bioinformatics 18: . https://doi.org/10.1186/s12859-017-1793...
  10. CRISPR Activation: A Practical Guide

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    Blog Post
    ...deliver the appropriate isoform(s) of a given gene, as, many times, the isoform(s) common to a particular...Tuttle is a medical student at Tufts University and former researcher at the Wyss Institute for Biologically...
  11. Five Popular Model Organisms, Part 2

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    Blog Post
    ...model in medical research with an archive of information and data that has yet to be produced with mice...Arabidopsis. There are also important tips for plant transformations and the use of Arabidopsis in the classroom...functional genomics study in bacteria with superior performance. Nature Communications, 9(1). https://doi.org...
  12. Harnessing Bacterial Toxins for Allelic Exchange

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    Blog Post
    ...rely on exogenous enzymes, require multiple transformation and plasmid curing steps, and can generate ... markers for allelic exchange. To do this, we performed the following modifications to pDS132 (Philippe...Hospital where he is now an attending physician. He performs post-doctoral research in the lab of Matthew Waldor...
  13. Fluorescent Proteins 101: When GFP lets you down

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    Blog Post
    ... GFP is such a well-validated, all-round good performing probe, it is the first choice when selecting ...Zacharias et al., 2002). GFP needs oxygen Chromophore formation requires a fully folded beta-barrel structure,...C-terminal fusions (Adjobo-Hermans, 2011). Structural information facilitates the choice of the proper site, although...
  14. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...generate non-phosphorylated recombinant protein and perform in vitro kinase reactions. Often, however, researchers... serves as an intermediate on the path to the formation of cysteinyl-tRNA (4). The discovery of a phosphoserine...PMC4383998. 3. Li, Tong, et al. "Cysteinyl‐tRNA formation: the last puzzle of aminoacyl‐tRNA synthesis."...
  15. A Control for All Seasons

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    Blog Post
    ...different types of controls and be able to use this information to design thoughtfully and thoroughly controlled...be due to natural biological variation. This information is critical for correctly interpreting experimental...experimental technique before starting. For more information about antibodies and antibody-based experiments...
  16. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...T7 promoter upstream of the left homology arm, perform in vitro transcription, and use the IVT product... and infundibulum. In vivo electroporation is performed immediately after injection. Image from Ohtsuka...contrast to standard microinjection, i-GONAD is performed directly on pregnant female mice using a small...
  17. Building Global Connections with the International Mentorship Program USA-EUROPE

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    Blog Post
    ...thought that it was necessary to develop a single platform to connect talented professionals (who migrated...nominations for government fellowships. The closing and formal evaluation of the pilot program IMP 2014/2015, ... success of this program. As we build a base of former mentees, we encourage them to get involved and ...
  18. Finding Your Science Policy Path

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    Blog Post
    ...societies, and industry through convening activities, formal lobbying, or strategy setting.  The “little p” ...failures, and challenges you’ve overcome. This information may help you determine what parts of your current...current job you want to keep or change and can inform your move into science policy. Read broadly about...
  19. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ... to consider how dyes can influence receptor conformation and signaling activity​​ (Sridharan et al., ... which is key to observing interactions and conformational changes in proteins. Both short distances of... expand the self-labeling capabilities of this platform. The Janelia Fluor ligands provide brighter and...
Showing: 601 - 620 of 963 results