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We narrowed to 639 results for: des.2

Showing: 501 - 520 of 639 results
  1. Plasmids 101: Golden Gate Cloning

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    Blog Post
    ...1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning Golden Gate cloning...customized to direct assembly of DNA fragments. When designed correctly, the recognition sites do not appear... scheme is as follows: the gene of interest is designed with Type IIS sites (such as BsaI or BbsI), that...and do not appear in the final construct. The destination vector contains sites with complementary overhangs...ligation can be done in one 30-minute reaction. The destination vector and entry vector(s) are placed in a single...IIS enzyme and ligase. Although the original destination vector + insert may spontaneously religate, this... be re-digested. In contrast, formation of the desired ligation product is irreversible because this construct...
  2. Plasmids for Endogenous Gene Tagging in Human Cells

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    Blog Post
    ... later this year. Designing the donor plasmid Figure 2: Donor plasmid design strategy. A donor ...with each cell line. The editing strategy and design described above can be used to create similar donor ...GFP tag and 1kb of homologous DNA flanking both sides (about 2.7kb total)—is inserted into a plasmid backbone...plasmid to prevent the plasmid from being cut and destroyed by CRISPR/Cas9 during transfection. Making the...use electroporation, a technique that briefly destabilizes the cell’s outer membrane and allows the components...Addgene’s CRISPR-based Protein Tagging collection includes systems for tagging mammalian cell lines, Drosophila...
  3. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

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    Blog Post
    ...still depolarize neurons at lower MOIs. Figure 2: ChRgers are delivered by intravenous injection into... model to help them construct “designer” channelrhodopsins with desired characteristics. The model was...Viviana Gradinaru’s lab used machine learning to design new channelrhodopsins that are more light sensitive...constrained and poorly understood. It’s hard to design new channelrhodopsins with high-throughput proteins...Using machine-learning to overcome channelrhodopsin design challenges To overcome these challenges, the Arnold...experiments. The sequences in this library were designed using structural information about parental channelrhodopsins...existing channelrhodopsins CoChR and/or ChR2(H134R). Designer channelrhodopsins paired with a systemic AAV allow...
  4. Hot Plasmids and Viral Preps - March 2021

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    Blog Post
    ...neuron-astrocyte, and cancer-stromal cell pairs.   Figure 2: Schematic of the interaction between the sender cell...repository that will help epigenetic researchers custom design their own engineering plasmid tools in about a ...cloning, the Haynes lab’s Protocols.io protocol describes an elegant system that allows for the assembly... cells that make up the human body and G-baToN provides versibility in tracking and modulating the interactions... inhibitor response 1 (TIR1) expressed under a desired promoter with a specific spatiotemporal expression...interests via CRISPR techniques. This tool-kit includes an array of FPs as well as additional useful tags...libraries encode target sites flanked by randomized nucleotides in place of a protospacer adjacent motif (PAM...
  5. Antibodies 101: Affinity Reagents

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    Blog Post
    ...Antibody Fragments. Antibodies (Basel). 2019 Apr 9;8(2):28. doi: 10.3390/antib8020028. PMID: 31544834; PMCID... as the OG (as the kids say) affinity reagents deserved to be celebrated, there are many other amazing...its thorns and every affinity reagent has its downsides. These reduced-in-size options often have reduced...°C and can refold correctly after denaturing.  Designed ankyrin repeat proteins, or DARPins, are genetically...a wide variety of targets, including proteins, peptides, amino acids, drugs, metal ions, and cells. They...
  6. Viral Vectors 101: Types of viruses

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    Blog Post
    ...Era of Personalized Medicine. Genes Dis. 2017 Jun;4(2):43-63. DOI: 10.1016/j.gendis.2017.04.001 Maetzig...about a specific virus? We have you covered with guides on lentivirus, retrovirus, adenovirus, and AAV....
  7. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

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    Blog Post
    ...improvement in muscle physiological performance.  2. Trans-splicing Overview: For trans-plicing, splice...doi.org/10.1002/emmm.201302948 Trapani I, Toriello E, de Simone S, Colella P, Iodice C, Polishchuk EV, Sommella...physical size of the genomes is still ~4.7kb. But despite this, larger functional transgene products were...HDR event can’t occur. Lastly, depending on its design, donor A may be capable of expressing a truncated...
  8. Antibodies 101: Multiplex Immunofluorescence

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    Blog Post
    ...signal separately for each target.   Figure 2: Examples of simultaneous multiplex IF labeling of... Brewer, M., McDonough, L., Zhu, Y., Neumann, E., De Caestecker, M., Gutierrez, D., & Spraggins, J. (2022...between the targets. Or, it might be sufficient to describe or classify the labeled structures visually; for...will vary depending on your specific experimental design. Like selecting an antibody, some of the requirements...
  9. Addgene: The First Twenty Years

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    Blog Post
    ...beginning Let’s take a quick flash back to 2004. “Shrek 2” was the most popular movie in theaters, the Razr ...business practices, like accepting e-signatures and designing a website users could easily navigate. This meant...Read the Addgene guide: https://www.addgene.org/guides/crispr/” — SirHamhands, responding to a request...
  10. Mouse Modeling, Part 1: Genetically Engineered Mice

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    Blog Post
    ...strain (which will be covered in more detail in part 2 of this blog series) controls the cell types in which...reporter gene, which replaces the targeted gene and provides a way to track successfully engineered cells. ...genotype each animal to identify the ones with the desired genotype. Inducible and conditional systems Conditional...
  11. How to Prepare for an Industry Interview

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    Blog Post
    ...company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer by looking at...method seems simple, I found it hard to remember to describe all four parts of a STAR answer once the interview... because sometimes companies have relaxed dress codes for interviews. You can also ask HR for clarification...
  12. Tips for Titering Your Lentiviral Preps

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    Blog Post
    ...protocols 4.4 (2009): 495-505. PubMed PMID: 19300443. 2. Geraerts, Martine, et al. "Comparison of lentiviral...virus will not only ensure that your infection is designed in the best possible way, but it may also save...the transgene itself. Many researchers prefer to design primers that target the common features of viral...
  13. 3D Printing Meets CRISPR Cas9

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    Blog Post
    ...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology,...BioMolecular Modeling and the CEO of 3-D Molecular Designs. Have you ever held your favorite protein in the...favorite protein in your hand? At 3D Molecular Designs, we create physical models of proteins using 3D...off a small, family-run business – 3D Molecular Designs – to commercialize some of the best instructional...reaction to this model on his blog. Or, for a description of how effective this model was in explaining... up close and personal, visit the 3D Molecular Designs web site. Or, if you are visiting a local classroom...check out our hands-on teaching tools that are designed to make the molecular world real for students....
  14. Open Resources and Plasmid Tools For Studying C. elegans

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    ...unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust GAL4...of 3’UTRs, the Arizona State University (ASU) Biodesign institute developed a 3’UTR database specifically...specifically for C. elegans. This database provides detailed information about C. elegans 3’UTRs structures, ...imaging processing. The combination of these methods provides unique opportunities to develop computational ...Addgene In addition to the wealth of open resources described above, there were many plasmid tools presented...self-excise, and covalently ligate their flanking peptides together (Wang et al., 2018).” With the split ...continuously curated as we continue to receive plasmids designed for the worm so check back frequently for what...
  15. CRISPR in the Clinic

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    ... Cure for HIV/AIDS. Int J Mol Sci. 2023 Jan 13;24(2):1563. doi: 10.3390/ijms24021563 Resources on Addgene.org...introducing a desired edit, but this particular class of treatments has quite the opposite aim: to destroy. One...potential for treatment is no longer an accurate descriptor; progress in clinical therapy is more fitting...One treatment currently in clinical trials includes Cas3 (the shredder member of the Cas family) as part...CRISPR therapies have made significant clinical strides since we first wrote this blog post. The future...
  16. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

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    ...C-terminus of human codon-optimized  cryptochrome 2 (CRY2) from Arabidopsis thaliana. When exposed to ...soCoChR-expressing system. Taken together, Shemesh et. al. describe a novel system for the precise mapping of neuronal...the single-cell level. Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid...information about spatial and temporal organization. The design use to detect PKA activity can be generalized to...choice for FRET assays where a dark acceptor is desired due to its very low quantum efficiency. ShadowY...
  17. RNA Extraction Without A Kit

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    ...156–159. https://doi.org/10.1016/0003-2697(87)90021-2 Chomczynski P, Sacchi N (2006) The single-step method...Allewell and Sarma, 1974). Additionally, ThermoFisher provides a protocol on how to integrate RNAlater® with ...
  18. Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)

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    ... adapted to a competitive format. Figure 2: Overview of using Competitive ELISA to quantitatively...ELISA is a modification of the sandwich ELISA as described here, the direct and indirect ELISA (more details...
Showing: 501 - 520 of 639 results