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  1. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

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    Blog Post
    ...endogenous retrovirus Rless glycoprotein (BaEVRless) had high rates of transduction for most cell types they tested...existing CRISPR delivery technologies, Nanoblades had higher rates for editing efficiency and lower rates of...zygotes, which results in albinism in mice. Of the eight mice born from Tyr Nanoblade treated zygotes, five...
  2. Mouse Modeling, Part 1: Genetically Engineered Mice

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    Blog Post
    .... In the first blog post in this series, I’ll highlight terminology you should be familiar with before...the use of long single-stranded DNA have led to a higher efficiency of homologous recombination for precise...
  3. Truncated gRNAs for Regulating Gene Expression

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    Blog Post
    ... of interest thus represents a straightforward means of exerting tight control over Cas9 nuclease activity...different genetic loci within a single cell. One might imagine transfecting cells with both the nuclease-positive...
  4. 3D Printing Meets CRISPR Cas9

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    Blog Post
    ...something silly like “See there, arginine 53 really is right there, at the end of helix 3…just like we said it...are useful. While a physical model of a protein might represent one aspect of the protein very well, it...while and you will wake up in the middle of the night dreaming about the conformational gymnastics that...
  5. Cloning Mammalian Cells with the Agarose Method

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    Blog Post
    ...colonies by (carefully) holding plates up to the light, where they appear as opaque dots (colonies should...the agarose onto the recipient plate at just the right temperature. If the agarose solution is too hot,...depending on the level of endogenous expression. We highly recommend the use of an electronic pipettor with...
  6. Polymerase Chain Reaction Overview and Applications

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    Blog Post
    ...primers.  Extension- The temperature is then raised slightly to Taq polymerase’s ideal temperature (70-75oC...which can be time consuming and challenging to get right. ddPCR uses a water-oil emulsion droplet technology...those stated above. Multiplex PCR is also used in high throughput SNP genotyping, gene deletion analysis...
  7. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

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    Blog Post
    ... Sometimes people don’t know how to start. They might want to use our promoters and terminators, but then...coding sequence to the MoClo system. It’s quite straightforward really and we’ll be producing an instructional...chances of doing something amazing will be equally high as for those who go to work in cancer biology. ...
  8. Tips for Using BLAST to Verify Plasmids

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    Blog Post
    ...to make the most of your time and sequence! You might also enjoy seeing how our quality control process...alignment results. The default setting is “Optimize for Highly similar sequences (megablast)”, which is very fast...nucleotide sequences, proteins often have repeated or highly homologous regions, which by default are ignored...
  9. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

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    Blog Post
    ...a collection of promises to not assert property rights for particular engineered genetic sequences; but...metabolite in a cell. The only way to do that at scale right now is to grow up a bunch of cells, sacrifice them...chemicals, to make more medicine. The parts we have right now are well adapted to work in prokaryotic bacteria...
  10. Plasmids 101: Secondary Nanobody Toolbox

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    Blog Post
    ...available Not commercially available Small but Mighty: Advantages of Nanobodies Although antibodies have...target antigen to about 2 nm, which can lead to higher resolution for super-resolution microscopy. Nanobodies... two questions, refer to this table to find the right nanobody for your work. Species Antibody Subclass...
  11. CRISPR 101: Ribonucleoprotein (RNP) Delivery

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    Blog Post
    ... off-target effects. Cas9 RNPs are detectable at high levels shortly after transfection, and are quickly...efficiency, while biolistic bombardment, which can have high efficiency, requires specialized equipment, such...microbiology.     References 1. Chen, Sean, et al. "Highly efficient mouse genome editing by CRISPR ribonucleoprotein...
  12. Course-Based Undergraduate Research Experiences (CUREs)

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    Blog Post
    ...trainees’ very way of thinking too. Albert Einstein might have put it best when he stated, “the value of an...pathways (precisely the type of exercise Einstein might have resented!). In 1997, Joseph Provost, then a...like an exciting opportunity for your classroom, I highly recommend you try implementing one. It's likely...
  13. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

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    Blog Post
    ... know the basic principles of IF, we’ll take a slightly deeper dive into those steps and provide you with...commonly used cross-linker for IF. However, you might come across a couple other terms that can cause ...especially if left on for too long or used at too high a concentration.      Figure 2: Examples...
  14. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

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    Blog Post
    ...A) Streamlined VchINT expression constructs. (B) High integration efficiencies achieved with VchINT, at...mini-transposon Transform plasmid(s) into cells and plate overnight Screen colonies phenotypically or by PCR/sequencing...Sternberg SH (2020) CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering...
  15. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ...standard curve for qPCR can be challenging to get right. Here at Addgene, we use a linearized plasmid standard...very few positive droplets. In the image to the right, there is approximately 1 positive copy per microliter...droplet generator to the PCR plate. Maintaining a high droplet count is important for calculating the concentration...
  16. Viral Vectors 101: Preparing Pooled Libraries

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    Blog Post
    ...depositing lab. (For our more popular libraries, you might receive a similarly-sized aliquot from an amplification...ll want to infect your cells with the virus–the right amount of virus, that is. Ideally, each cell will...Gootenberg, J. S., Abudayyeh, O. O., Platt, R. J., Brigham, M. D., Sanjana, N. E., & Zhang, F. (2017). Genome-scale...
  17. Using Video to Share Your Science: We Share Science

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    Blog Post
    ...interested audiences quickly and concisely – video might be part of the answer. Problems with written publications...TedTalks are very popular, though they focus mostly on highly innovative ideas rather than on the day-to-day ...that provides links to the videos, keeping the copyright and control over the videos with the researcher...
  18. The PAM Requirement and Expanding CRISPR Beyond SpCas9

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    Blog Post
    ...particular gene. Furthermore, a target sequence may have high homology elsewhere in the genome. These off-target...Sontheimer EJ (2018) NmeCas9 is an intrinsically high-fidelity genome-editing platform. Genome Biol 19...Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 556:57–63 . https://doi....
  19. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

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    ... #MolluscMonday: #Crepidula atrasolea juvenile, light microscope with polarizing filter. Development is...an extreme number of nerve cells and an unusually high level of RNA recoding in the brain. RNA editing ...DNA in the eggs to get transgenic animals. They might be immortal. Manihot esculenta (Blake Meyers) Cassava...
  20. Designing Your Chalk Talk for the Academic Job Interview

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    Blog Post
    ...million dollar sales pitch. Given this large sum, one might expect that applicants would work for months to ...research? This is the central idea. It should be at a high level, broadly understandable to scientists outside...engaged by your presentation, then you’re on the right track. Furthermore, practice getting back to your...
Showing: 701 - 720 of 995 results