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We narrowed to 450 results for: abo.5

Showing: 121 - 140 of 450 results

  1. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    Blog Post
    ...   The Mello lab also reported that modifying the 5’ modification of oligos used to generate linear dsDNA... animal.  PAM sequence considerations For changes 5’ to the PAM, sequences corresponding to the same strand... using two Cas9 targets. An orientation where the 5’ ends of the PAMs are facing outwards, ensuring the...Edraki A, Watts JK, Sontheimer EJ, Mello CC (2018) 5′ Modifications Improve Potency and Efficacy of DNA...learn more about genome editing with CRISPRs? Check-out Addgene's CRISPR Resources Learn about gRNA design...interest, I encountered low efficiency (0.13%) and laborious handling (screening 768 F1 animals). Recovering...

  2. Selecting Your Plasmid Purification Kit

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    Blog Post
    ...of culture Approximate DNA yield** Miniprep 1–5 mL 5–50 µg Midiprep 25–100 mL 20–200 µg Maxiprep...for example. These kits are designed to handle ~1–5 mL of liquid culture. Using a higher volume can affect...µg Megaprep 0.5–2.5 L 1.5–2.5 mg Gigaprep 2.5–5 L 7.5–10 mg *Can vary from kit to kit, check the...columns we’ve been talking about!) are a significant source of waste in laboratories. To help combat this, ...dependent on many different factors as we detailed above, so be sure to pick what kit works best for you!...

  3. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

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    Blog Post
    ...Cellular Biology 5(11): 3274–79. PubMed PMID: 2427927. PubMed Central PMCID: PMC369144. 5. Kranz, Andrea...Cellular Level in the Mouse.” Nature Biotechnology 21(5): 562–65. PubMed PMID: 12665802. 9. Villa, Katherine... on the Addgene Blog Learn More about FLEX-vectors Learn More about Cre-Lox Check out Retrograde AAVs...expression have been developed. Read on to learn more about FLP-FRT, Cre-lox, and how combinations of FLP and...

  4. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

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    Blog Post
    ...their Roller phenotype. If SEC is inserted at the 5’ end of a gene, the SEC separates the targeted gene...are then heat shocked to remove the SEC and after 5-6 days of growth, wild-type animals that have undergone...just one injection step! The entire protocol takes about 2-3 weeks but requires less than eight hours worth... this species, namely, the robust HDR mentioned above, an accessible germline, and multiple selectable...modular system that requires very little hands-on labor and could facilitate the tagging of every protein...

  5. COVID-19 Resources

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    Collection
    ...protein, activating its membrane fusion function.( 5 ) TACE - (ADAM17) a metalloprotease that cleaves hACE2...lightning speed to elucidate as much as they can about this novel virus. It is now known that SARS-CoV-...and open-access SARS-CoV-2 detection assay for laboratory and home testing. Kellner, et al. bioRxiv 2020.06.23.166397...

  6. Antibodies 101: Epitope Tags

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    Blog Post
    ...the protein with enterokinase, which recognizes the 5 amino acids on the C terminal of the tag. Of the most... most common tags, FLAG is the most charged, with 5 negatively charged and 2 positively charged amino ...Tags have a lot of major benefits – as discussed above – but there are a few instances when they should... hindrances, and destabilization can be brought about by the tag so it’s important to validate tagged ...

  7. New and Upcoming Viral Vectors - June 2019

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    Blog Post
    ...AAV9, AAVrg pAAV-hSyn-DIO-mCherry 37825 AAV1, 2, 5, 8, 9 (20 µL size serotype testing) pAAV-CAG-GFP ...size serotype testing) pAAV-CAG-GFP 50465 AAV1, 2, 5, rg (20 µL size serotype testing) pAAV-hSyn-EGFP ...vector name and serotype you’d like to be notified about. Controls Plasmid Serotype Name 37825  AAVrg...vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols and...

  8. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

    Type
    Blog Post
    ...expected to be >12 kb. However if it recombines at the 5’ and 3’ LTR regions, it splits into two smaller plasmids...recombined vector backbone.   Ready, set, grow! Our 5 Tips for Troubleshooting Viral Transductions will ...molecular biology and viral vector protocols Learn about the common lab E. coli strains Find tips for viral...

  9. Visualizing Translation at the Single Molecule Level

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    Blog Post
    ... codons/second. They also explored the effects of 5’ UTR variants in translation of cell cycle protein...Emi1, showing that ~80% of transcripts with a long 5’ UTR were translationally silent. Both of these methods...Additional Resources the Addgene Blog Learn More about SunTag Catch up on Your GFP Background Choose the...

  10. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...Coli Synthetic Biology.” ACS Synthetic Biology, vol. 5, no. 10, American Chemical Society (ACS), 2 May 2016...Traditional Biology.” ACS Synthetic Biology, vol. 5, no. 1, American Chemical Society (ACS), 4 Nov. 2015....” PLOS ONE, edited by Alvaro Galli, vol. 13, no. 5, Public Library of Science (PLoS), 30 May 2018, p.... vol. 8, no. 11, American Chemical Society (ACS), 5 Nov. 2019, pp. 2593–606. Crossref, https://doi.org... constructs that compose genetic circuits and metabolic pathways. For many MoClo systems, the individual...interested in recombinant protein purification and metabolic engineering. If instead you want to build genetic...designed for flexibility. The Fragmid kit described above works in both mammalian systems and Drosophila. ...

  11. CRISPR-mediated Plant Base Editors

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    Blog Post
    ...the success of precise change via HDR limited to 0-5%. This blog post covers a brief review of different...SpCas9 rAPOBEC1 NGG 6 SpCas9 Target-AID (PmCDA1) NGG 5 SpCas9-VQR rAPOBEC1 NGA 8 SaCas9-KKH rAPOBEC1 ...PMID: 25733849. PubMed Central PMCID: PMC4371917. 5. Shimatani, Zenpei, et al. "Targeted base editing ...gene, Xw, except for a single base. If you heard about CRISPR, one question probably comes to mind: can...compatibility All of the base editors mentioned above are SpCas9 nickase-based and depend on the presence... HDR methods for rice crop improvement in the laboratory of Yinong Yang at PSU.   References 1. Komor,...The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018). Additional resources on the Addgene blog...

  12. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

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    Blog Post
    ...methodology with T cell receptor engineering (Ref 5)  as described by Blue Flame Addgene depositor Professor...PMID: 26343581. PubMed Central PMCID: PMC4597191. 5. Eyquem, Justin, et al. "Targeting a CAR to the TRAC...Cancer, Inflammation and Immunity and learn more about “... recent findings in basic immunology, cancer-immune...curing cancer in cross-disciplinary projects and collaborations. Summarized by Professor Michael Karin from...This was followed by a talk from Professor Karin about alcohol and obesity as driving factors for chronic...got a chance to talk with Dr. Friedmann-Morvinski about tools used to analyse the tumor microenvironment...community, contact [email protected] to find out more about our free deposit and distribution service! References...

  13. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...phosphoserine directly into recombinant proteins (5). Optimization of this system defined three components...into recombinant proteins, such as the kinase MEK1 (5). MEK1 is inactive without requisite dual phosphorylation...307.5717 (2005): 1969-1972. PubMed PMID: 15790858. 5. Park, Hee-Sung, et al. "Expanding the genetic code... one of the MEK1 phosphomimetic residues in the above study with phosphoserine (to create a pSer/phosphomimetic...non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid - Addgene...interested in the intersection of cell signaling, metabolism, and the mighty mitochondrion. You can follow...Blog Verify Your Constructs with Colony PCR Read about Tags for Protein Purification Resources on Addgene.org...

  14. Plasmids 101: Restriction Cloning

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    Blog Post
    ...downstream of the promoter, you can add an EcoRI site just 5’ of the start of the gene and a HindIII site just ...two HindIII digested ends will anneal leaving the 5’ end of your gene just downstream of the promoter ...plasmid : insert ratio” in the ligation reaction. 5. Purify the finished plasmid   Once it looks like... (post or pre run in the table above, respectively). Some of the above stains require you to visualize...equivalent to backbone + insert, unsuccessful 2 and 3 above), you can try ligating the recipient plasmid alone...get experimenting!   Additional Resources: Read about Gel Stains on the BitesizeBio Blog  Additional ...

  15. Pushing the Limits of DNA Assembly

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    Blog Post
    ...makes it so forgiving, is the incorporation of a 3’-5’ exonuclease in the reaction mixture. The exonuclease...expression cassettes for yeast transformation can take 5 days. NEBuilder shortens that time to one day by simplifying...scarless plasmid inserts.  Want to Learn More about DNA Assembly? Check Out Our Plasmids 101 eBook! ...to produce sgRNA for CRISPR experiments can be laborious, requiring multiple cloning steps. You can simplify...Additional Resources on the Addgene Blog Learn More about Gateway Cloning Gibson Assembly and Other Long-Homology...

  16. Evolution of Lab Techniques

    Type
    Blog Post
    ...restriction enzymes that cut DNA at specific sites (5). Imagine that you have a genome with unknown sequence... allowed for many seminal scientific discoveries (5, 8). While newer techniques have largely replaced ...PMID: 19283728. PubMed Central PMCID: PMC2743315. 5. Roberts, Richard J. "How restriction enzymes became...abundant in a sample. Whereas Southern blotting is labor intensive and requires a large amount of high-quality...DNA technologies that are now commonplace in laboratories. As scientific technologies rapidly progress...PMC4411503. Resources on the Addgene Blog Learn More about Restriction Cloning Get Tips on Analyzing DNA Sequencing...

  18. A Needle in a Base-Stack: Cas9 Structural Biology

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    Blog Post
    ...recognizes the PAM sequence 5’-NGG-3’, where N can be any base. In Figure 5, we can see why: two arginines...complementary strand is mismatched.   Figure 5: S. pyogenes Cas9 bound to guide RNA and target DNA... Complex with Guide RNA and Target DNA. Cell, 156(5), 935–949. https://doi.org/10.1016/j.cell.2014.02.001...nascent heteroduplex and abolish cleavage (Pattanayak et al., 2013). As mentioned above, however, Cas9 sometimes..., 2022), indicating that there is more to learn about this process!      Figure 3: Crystal structure...rest of Cas9 and is highly flexible; it can rotate about 140 degrees between inactive and active conformations...conformational changes that make Cas9 such a valuable laboratory tool are still an area of active research. New...

  19. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...Zhang et al. designed multiple gRNAs to target the 5’ long terminal repeat (LTR) of the HIV-1 genome, which...Kaminski et al. designed gRNAs to target the HIV-1 5’ and 3’ LTRs and expressed them along with Cas9 in...Replication and Accelerate Viral Escape. Cell Rep. 2016 Apr 5. PubMed PMID: 27068471. Ramanan V, Shlomai A, Cox ...reservoirs, chronic inflammation, and negative metabolic effects of ART. Why can’t the body mount an immune...

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

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    Blog Post
    ...provides both the endonuclease and ligase function while 5’ adenosine overhangs left by Taq perfectly anneal ...articles; so if you are interested in learning more about any one of them, please click on the links throughout...laying around the lab. If not, you’ll be looking at about $50-$60 per enzyme. Once you’ve got a few of these... relatively inexpensive and can be obtained for about $70, resulting in a price of a few dollars per reaction...
Showing: 121 - 140 of 450 results