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We narrowed to 634 results for: des.2

Showing: 481 - 500 of 634 results

  2. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ... enrich the microvesicles containing AAVs (Figure 2). Since the development of the exo-AAV isolation protocol...producer cells and purification protocols often describe a process that begins with cell lysis to release...

  3. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    ...and open a new conversation about GMOs.  Challenge #2: Technical obstacles to plant synthetic biology In...have been a model organism for plant research for decades, and the biochemical pathways involved in central...

  4. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. PubMed. Find the plasmids from this publication ...cleaved precisely as predicted with no addition of nucleotides to the 5’ spacers. The 3’ ends were modified ...advantage over other methods that favor certain nucleotides at the 5’ end. PTG construct schematics. Scissors...the plasmids made by Xie et. al are specifically designed for plant expression, PTGs will likely be adaptable...worries, I've summarized the three strategies described in this post in the graphic below - including ...

  5. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ... level.     Fig. 2 Tetracycline on/off promoters and modes of action Check out Addgene's ...and SMASh degron tagging vector! Destabilizing domains Destabilizing domains (DD) are akin to degron ...post-translational level, unlike most of the other mechanisms described. Fig. 3 dTag mechanism of protein degradation...

  6. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...extraction solution, runs the protocol, then uses around 2–4 µl in each 50 µl PCR reaction. The gDNA extracted... early onset Parkinson Point mutations. Cell, 146(2), 318–331. https://doi.org/10.1016/j.cell.2011.06.019...template vector design guidelines? A7: A few notes below are considerations for designing HR donor. Generally...usually pick 3–6 guides around the region to find the most efficient guide (most guides would work, but... this ‘split RNA’ design is not as efficient in genome editing as the second design (in pX330/PX335), ... CRISPR plasmid tools and numerous experimental design decisions to make, it makes sense that scientists...CRISPR forum's most popular questions and answers. Designing your CRISPR genome editing experiment Q1: Should...

  7. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    ...closely mimicking the future application. In figure 2 we provide an example of the comparison of the practical...tables with fluorescent protein properties and provides a quick way to compare fluorescent proteins, e.g...been used in yeast by Lee et al (2013) and in nematodes by El Mouridi et al (2017) and Heppert et al (...practical brightness. Therefore, practical brightness provides a better picture of what can be expected in a ...

  8. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...recombineering (2) to simultaneously incorporate multiple single-strand DNA (ssDNA) oligonucleotides (oligos)...backbone, which expresses the gRNA from a Drosophila U6:2 promoter and Cas9 from the actin 5C promoter. Addgene...backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors then used GMAP assembly...number of tandemly expressed gRNAs respectively) and 2) the size of the deletion. Smaller deletions generally...relaxed gRNA design parameters. To demonstrate the effectiveness of their library design, Moffat laboratory... pCAG-T7-TALEN(Sangamo)-Destination series, pCAG-Golden-Gate-Esp3I-Destination Takashi Yamamoto pcDNA-TAL-NC2...vector, and besides being used for restoring strain prototrophy, can also aid in designing self-establishing...

  9. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ..., but at a lower efficiency (~20%). Figure 2: Schematic of multicomponent assembly with SLIC. Multiple..., the use of LIC is often limited to specially designed plasmids. Sequence- and ligation-independent cloning...the “damage” and producing a complete plasmid. Besides being compatible with any vector, the SLIC protocol...

  10. Antibodies 101: Isotypes

    Type
    Blog Post
    ...IgM, IgD, and IgG are deleted.     Figure 2: Isotype switching requires DNA recombination of the...molecules are recruited by the antibody to help destroy and remove a pathogen. Different isotypes also ...well as the genes that encode those proteins are designated by the lower-case Greek letters μ, δ, γ, ε, and...

  11. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...available in the adenovirus section below!   Fig. 2 - Viral vector components of AAV Adenovirus Adenoviruses...promoter should flank this component as well. On both sides of this promoter-gene combo are two long terminal...Laboratory Press; 1997. Principles of Retroviral Vector Design. Available from: https://www.ncbi.nlm.nih.gov/books...

  12. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...cellular responses in addition to antibody response 2) very high immunogenicity 3) and long-lasting immune...biology – they can deliver materials to specific destinations, haul a variety of cargo, and even handle a ...delivery of the actual virus to which immunity is desired. Instead, components of the virus are now delivered...

  13. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...PubMed PMID: 8491198. PubMed Central PMCID: PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human...target cells and organisms, and when that cargo is designed to make you sick…well, you feel it. But the same...host genome or temporarily express whatever it encodes. This ability to infect cells with a high efficiency...

  14. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    ...JS, Legube G, Miller KM Genes Dev. 2015 Jan 15;29(2):197-211. DNA Cross-Bridging Shapes a Single Nucleus... articles reporting chromatin-related research describe the production of plasmids that might be useful...colleagues and guest bloggers have written posts describing new methods and tools for studying chromatin ...

  15. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...higher quenching of mScarlet emission. Figure 2: Ace-mScarlet is a fusion between the voltage-sensitive...showing the potential for broad use. This kit is designed for use with acceptor vectors from the MoClo Toolkit...tracing Article contributed by Erin Sanders CreLite, designed by the Eisenhoffer lab, is a new tool that increases...

  16. Plasmids 101: Cre-lox

    Type
    Blog Post
    ..., N. and Hamilton, D. 1981. PubMed PMID: 6276558. 2. Site-specific DNA recombination in mammalian cells...using a resistance marker; however, it is often desirable to remove the marker after the initial selection... Combining this with some of the loxP methods described above, a genetic modification can be restricted...

  17. Plasmids 101: Golden Gate Cloning

    Type
    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...

  18. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    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...

  19. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

    Type
    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...

  20. Hot Plasmids and Viral Preps - March 2021

    Type
    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...
Showing: 481 - 500 of 634 results