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We narrowed to 778 results for: Dos

Showing: 391 - 420 of 778 results
  1. Antibodies 101: Fab Fragments

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    Blog Post
    ...Imagine an antibody. Do you immediately visualize a Y-shaped protein reminiscent of the Addgene mascot...sure to think about what pieces of the antibody you do and don’t need, and how the protein will interact...
  2. Antibodies 101: Stripping and Reprobing Western Blots

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    Blog Post
    ...during your western and your results look… funky? Do you need to start a whole new blot? Never fear: membrane...directly to buffer without a re-wetting step. Important: do not dry your membrane with antibodies on it if you...
  3. Viral Vectors 101: Virus Safety

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    Blog Post
    ...virus to mice, part of your preparation should be doing your best to be well-rested and alert the day of...procedure. Mice move much more quickly than cell cultures do, and tired researchers are accident-prone researchers...
  4. Viral Vectors 101: An Introduction to AAV

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    Blog Post
    ... cell and animal models. AAVs are non-pathogenic, do not integrate into the host genome, and can be specifically...several years (Wonjo et al., 2013). Remember that AAVs do not integrate into the host genome and therefore ...
  5. Tips for Titering Your Lentiviral Preps

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    Blog Post
    ... all have their benefits and drawbacks. Labs that do not routinely produce virus may want to choose more...Additional Resources on Addgene.org Learn How We Do Viral Production at Addgene Read Our Lentiviral Guide...
  6. Transferable Skills Guide: Cross-team Communication

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    Blog Post
    ... only problematic word. How many different things do you simply refer to as a “tube”? We took steps towards...lab or collaborators will not have the context you do, which means you’ll need to provide that context ...
  7. CRISPRainbow and Genome Visualization

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    Blog Post
    ...ability to target almost every genomic locus, and to do so in vivo. Catalytically dead Cas9 (dCas9) cannot...CRISPRainbow requires very short, 11-mer gRNAs, which do not induce genome editing. If used with catalytically...
  8. Plasmids 101: Golden Gate Cloning

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    Blog Post
    ...fragments. When designed correctly, the recognition sites do not appear in the final construct, allowing for precise...these sites are eliminated by digestion/ligation and do not appear in the final construct. The destination...
  9. How to Design Your gRNA for CRISPR Genome Editing

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    Blog Post
    ...use, of course, depends on what you are trying to do – there’s no “best” tool among them. While this seems...gRNA depends an awful lot on what you are trying to do: gene knockout, a specific base edit, or modulation...importance in design – an optimized sequence will do little if it is in the wrong place, but because the...
  10. Important Considerations When Using AAVs

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    Blog Post
    ...from Addgene 1. Location, location, location! What do buying real estate and using AAVs have in common?...each serotype can transduce multiple tissues. So how do you choose the right serotype? Keep asking questions...injection. Examining serotypes 1, 8 and 9, not only do they undergo retrograde and anterograde transport... transport can be reduced by injecting a smaller dose of AAV and using a serotype with weak CNS penetrance...
  11. Plasmids 101: Using Transposons in the Lab

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    Blog Post
    ... human genome (Munoz-Lopez et al., 2010).  So why do transposons matter? The outcome really depends on...how retroviruses, like HIV, replicate. Class I TEs do not encode a transposase enzyme.  Class I TEs are...TEs are also known as DNA transposons, since they do not use an RNA intermediary when they move. Most ...
  12. Progress Towards a PAM-Free CRISPR

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    Blog Post
    ...non-coding RNA, cutting most anywhere within the RNA will do. Similarly, if you want to knock out a gene, a frameshift...introduced anywhere within the early coding region will do the job. What matters for these experiments is cutting...
  13. Technical Design of a Western Blot

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    Blog Post
    ...detection range of your protein. The most common way to do this is via a Bradford assay or BCA assay. Be sure...preserve specific protein modifications, we recommend doing a little more research into gel chemistry and/or...wide dynamic range for fluorescent proteins. They do tend to have increased background with higher exposure...your blot to calculate the amount in your sample. (Do ensure your blot is optimized so your standard curve...
  14. Antibodies 101: Designing Your First Flow Panel

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    Blog Post
    ...fluorescence when excited with ultraviolet (UV) light, they do so via aromatic amino acids found in all proteins...photons excite electrons of a fluorophore, they can do so at a range of wavelengths, rather than at just...1, emission spectra of neighboring colors can and do overlap. For example, APC overlaps noticeably with...
  15. When Fidelity Matters: A frank discussion about ligase fidelity

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    Blog Post
    ...biochemistry researcher at New England Biolabs. When do you need a high fidelity ligase—and when is an alternative...base pairing through minor grove contacts, and thus do not read specific base sequences, but are sensitive...differential is not known for certain, but may have to do with the slight melting of the 5 ́-terminus during...
  16. 10 Basic tips for mammalian cell culture

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    Blog Post
    ...use. This will prevent spills, but, if any spills do occur, they should be wiped with 70% ethanol immediately...morphologies, being familiar with these morphologies and doing a quick check under the light microscope will help...
  17. AAV Vector Quality Control: Going the Extra Mile with NGS

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    Blog Post
    ...they are present at a very low concentration. So how do we make sure to catch all of these potential mishaps...expected hits, we always get hits to “random” genes. Do these hits always mean the sample is contaminated...users titrate their AAV vectors to find the optimal dose that allows for sufficient transgene expression ...
  18. Of Myc and Men

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    Blog Post
    ...Do you ever wonder about the origins of some of the common techniques or tools you use in the lab? Take... to be less sensitive to sequence context.    So, do Schüchner, et al.’s results mean the end of 9E10’...
Showing: 391 - 420 of 778 results