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We narrowed to 282 results for: c rel

Showing: 181 - 200 of 282 results
  1. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ...construct. Reactions are often cycled between 37°C and 16°C to provide optimal reaction temps for both the...practices for assemblies of more than 10 modules often rely on two-step hierarchical approaches using different...pre-determined manner. If the original insert religates into its original vector, it will subsequently...
  2. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    Published
    Jan. 16, 2020, 2:18 p.m.
    ...calculate the viral titer: GC/mL = {[(R*C)(1000/V)]*D} R = Reaction volume C = Copies/uL V = Volume of virus ...polymerase chain reaction Browse our blog posts related to viral vector protocols and tips Read all of ... by qPCR Browse our collection of viral vector related protocols Learn more about the Addgene viral service...
  3. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...Most FP fusions for FRET studies are at the N- or C-terminus of your target protein, but even these modifications...can insert your protein of interest, resulting in a C-terminal fusion of a tetracysteine tag. In addition...for observed FRET efficiencies between 80% and 10%. C Cy3NTA and Cy5NTA bind His tags similarly but with...poly-histidine tag often used for protein purification is a reliable FRET acceptor labeling site when used in conjunction...
  4. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ... epitope such as c-Myc, GFP or V5 can be used to tag the protein of interest on the C- or N-terminus. ...conformations and interactions. For difficult to release proteins, including those in the nucleus, more ...
  5. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ...into an attL-containing entry vector.   Method C: Restriction cloning of a restriction enzyme fragment...attL-containing entry vector.   Figure 5: Method C to create an entry clone: Restriction cloning the ...their desired DNA sequences. Gateway technology relies on the two reactions described below: The BP Reaction...
  6. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...overhangs Cas13a (C2c2) 2x HEPN crRNA RNA 3' A, U, or C (not required by all orthologs) Specific RNA cleavage...spacer sequence and consists of a single A, U, or C base pair. In bacteria, once Cas13a has recognized...Class 2, candidate 1, 2, or 3). C2c1 and C2c3 are related to Cpf1, but they require both a tracrRNA and crRNA...
  7. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    Published
    July 15, 2025, 1:15 p.m.
    ...reactions, where a fraction of one of the four bases (A, C, T, or G) is supplied as a dideoxyribonucleoside triphosphate...regions of interest, or complex sequences such as G-C rich areas, secondary structures, or repetitive regions...widely used DNA sequencing technology. The process relies on “sequencing-by-synthesis,” where each base is...sequencing.  Prior to short-read sequencing, Addgene relied on Sanger sequencing for QC. We occasionally still...Second-generation sequencing”)  Short-read sequencing also relies on sequencing by synthesis, but modifies the original...
  8. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    Published
    Aug. 10, 2021, 1:15 p.m.
    ...agarose gel to confirm product presence, size, and relative quantity. End point PCR is most commonly used ...     References and Resources References Shen C-H (2019) Amplification of Nucleic Acids. In: Diagnostic...reagents and several heating and cooling steps. PCR relies on heat resistant DNA polymerase from the thermophilic...
  9. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...8.2 kb, and in vivo transgene expression ceases relatively quickly due to immune response against AdV. Importantly...4. Mittereder, Nanette, Keith L. March, and Bruce C. Trapnell. "Evaluation of the concentration and bioactivity...
  10. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    Published
    June 12, 2014, 3:30 p.m.
    ...Nicotiana benthamiana. N. benthamiana is a tobacco relative and a good model for plant disease because it ...assembly of multigene constructs. Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. PLoS One. 2011...tools work in any plant? Patron: The MoClo kit is entirely for dicots, not grasses and wheat, but we do work... have your parts. That was the motivation for releasing a large parts kit. Once you have so many parts...
  11. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ...specificity of GFP expression for the indicated ssAAV5s. C) Quantifications showing high microglia transduction... microglia-targeting AAV plasmids here! Serrano, C., et al. (2023). Simple and Highly Specific Targeting...fast-spiking neurons. The same A-to-D mutation in the related voltage indicator Ace2N-mNeon also led to similar...dCas9-VPR, prime editing, and delivery of disease-relevant large proteins. They demonstrated REVeRT in various...
  12. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    Published
    Dec. 2, 2019, 2 p.m.
    ...these methods a try.  References:  Branon, Tess C., et al. "Efficient proximity labeling in living cells...1837. PubMed PMID: 30676735. Kim, D. I., Jensen, S. C., Noble, K. A., KC, B., Roux, K. H., Motamedchaboki...array of biological contexts.  Proximity labelling relies on the labelling of neighboring proteins within...interactions in vivo (Ge et al., 2019). This technique relies on an S. auerus transpeptidase sortase A variant...
  13. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ...groups making use of the new optogenetic tools in C. elegans and in Plasmodium. For those who do want ...those optogenetic controls, gene expression levels reliably increased in vitro by a factor of 10 to 20. A ...modifications in the genome,” Zhang said in MIT’s press release. In an accompanying Nature News & Views Andreas...
  14. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    Published
    June 4, 2024, 1:15 p.m.
    ...similar nuclease folds found in other proteins. The C-terminal domain (CTD) of the protein also belongs ...https://doi.org/10.1038/s41586-022-04470-1 Cofsky, J. C., Soczek, K. M., Knott, G. J., Nogales, E., & Doudna...Sternberg, S. H., Redding, S., Jinek, M., Greene, E. C., & Doudna, J. A. (2014). DNA interrogation by the...the enzyme needs cellular recycling factors to release its handiwork (Sternberg et al., 2014). New discoveries...
  15. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    Published
    July 8, 2025, 1:15 p.m.
    ...; CDSns, coding sequence with no stop codon; TAG, C-terminal tag; TER, terminator. Created with BioRender.com... is very important that these stay frozen at -80 °C, since multiple rounds of freezing and thawing will...in what position do you want your gene to be in relation to the rest of the genes? And do you want it to...
  16. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ...solubility (Monnier et al., 2013). The linker keeps the C-terminus of one variable domain and the N-terminus...10.3390/antib2020193  Khantasup K, Chantima W, Sangma C, Poomputsa K, Dharakul T (2015) Design and Generation...to avoid animal use by using semi-synthetic or entirely synthetic VL and VH sequences. Phage display also...scFvs are also used for eye diseases, such as age-related macular degeneration (AMD) since direction injection...of the CAR facilitates signal transduction and release of cytotoxic granules from the T cell. When a CAR... CAR has an scFv that binds cancer-related antigens, binding of the scFv part of the receptor can activate...
  17. How to Write a Scientific Review Article

    Type
    Blog Post
    Published
    Feb. 16, 2017, 3:30 p.m.
    ... all the proteins in the class that showed the relative sizes and the conserved domains. This information...perfect. Good luck! References 1. Haery, Leila, Ryan C. Thompson, and Thomas D. Gilmore. "Histone acetyltransferases... maybe you’re researching a topic for which no relevant or recent review exists. Once you pick a topic...format also has the benefit of exposing you to relevant researchers in the field. Finally, you can sound...from start to finish. Find the information that’s relevant to you, note it, and move on. At the beginning...
  18. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ... deliver a large amount of DNA (~8 kb) with a relatively low immune response, it should come as no surprise..., Andre K, Stein D, Blick G, Greenberg RN, Kinder C, Zolopa A, Binder-Scholl G, Tebas P, June CH, Humeau... co-transfection of these plasmids, 293T cells released transgene-containing lentiviral particles into...
Showing: 181 - 200 of 282 results