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We narrowed to 950 results for: omp

Showing: 541 - 560 of 950 results

  1. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ... Figure 2: MoClo assemblies depend on compatibility of four-base overhangs flanking each genetic...To combat this, you can increase the volume of competent cells you plate after each transformation and/...high-throughput, we recommend transforming into chemically competent cells in a 96-well PCR plate. Always confirm your... MoClo user, but beginners or those performing complex assemblies should screen a minimum of 8–10 colonies...

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

    Type
    Blog Post
    ...interior target sites. In a second step, the two compatible target sites are cleaved out, leaving behind ...a vector with the inverted DNA flanked by non-compatible target sites (Schnütgen et al. 2003). Flex vectors...within neuronal networks in living animals. To accomplish such cell specific manipulations in vivo, FLEx...knockout. In this experiment it is possible to compare neuronal morphology and synaptic connectivity before...

  3. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ... and the complexity of effectively deploying them in the lab somewhat mirrors the complexity of understanding.... Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult both to..., and jGCaMP8 viral preps distributed per year compared to other AAVs preps from 2016–2023. B) Inset showing...

  4. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...virus as self-complementary AAV (scAAV). However, scAAV vectors have limited size compared to single-strand...-multi (Walton et al. 2024). This platform is compatible with enrichment, single-cell sequencing, and ...virus’s most abundant proteins and a critical component for viral genome packaging. Addgene recommends...

  5. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...Host range depends on whether the host has the components to recognize the ori and allow for replication...organisms. pBBR1-based plasmids and plasmids in incompatibility groups N, P, Q, and W are known to have replicons...list of the bacterial species these plasmids are compatible with.  Table 1: Non-exhaustive list of common... and Srivastava 2013. Family Replicon Compatible with Shown to work in IncN pCU1 Gram-negative...

  6. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...reaction creates an expression clone with all of the components necessary for gene expression.     The LR... are performed, the next step is to transform competent E. coli cells and select the positive clones. ...functional. Advantages of the Gateway cloning method Compatibility and flexibility Once you generate the entry ...have generated and deposited their own Gateway-compatible plasmids with Addgene. These can be used to express...

  7. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...can be difficult to construct and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used to...Housden BE, Perrimon N, Collins JJ, Church G (2016) Comparison of Cas9 activators in multiple species. Nat Methods...transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https://doi.org/10.1038/nature14136...Kadoch C (2018) Interrogation of Mammalian Protein Complex Structure, Function, and Membership Using Genome-Scale...

  8. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...endogenous homologous recombination enzymes can also accomplish the desired genomic modification. This technique... which results in a merodiploid. This can be accomplished by selecting for an antibiotic resistance marker...scarless modification of bacterial genomes. It can accomplish large deletions, insertions, and even precise...PMC4513841. Latynski US, Valentovich LN. "DNA tuner: a computer program for the construction of polylinker sequences...

  9. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ... proteins in living cells and organisms. Both components of the chimera are encoded by DNA. Since researchers...When both orientations are available, a direct comparison of these fusion proteins may reveal which of ...the fusion protein can be verified and it can be compared to what is expected based on immunofluorescence...interactions) can be challenging. If a fusion can complement a knock-down or knock-out of the corresponding...

  10. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...to the popularity of GFP including (i) fast and complete maturation to functional, fluorescent protein ...diameter of about 2.4 nm (Hink et al., 2000). The complete beta-barrel is necessary for its fluorescence ...genetically encoded tag and exogenously added components. For a review see Lotze et al. (2016). To date...is used to excite GFP, may also excite several components that are naturally present in cells or media....

  11. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...method for knockouts is Tracking of Indels by Decomposition (TIDE) (Brinkman et al., NAR). This technique...trace files of WT and edited cell lines. Then decomposition analysis is performed yielding a graph of edits...This method requires a set of control cells to compare the sequencing reads from your edited sample to...assessment of genome editing by sequence trace decomposition. (2014) Nucleic Acids Research Volume 42, Issue...

  12. The Advances Behind The World's First Personalized CRISPR Treatment

    Type
    Blog Post
    ...between research hospitals, universities, private companies, and federal regulators. But the speed is impressive...which could be repaired with an A→G edit on the complementary strand. But the invention of base editing doesn...approval, as well. If the editor and delivery components could be rigorously safety tested and reused,...adenine base editor with improved Cas domain compatibility and activity. Nature Biotechnology, 38(7), 883...

  13. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...the production of lentiviral vectors that are competent for one round of infection (AKA transduction in...progeny are produced (Figure 1).  Figure 1: Comparison of the life cycle of a lentiviral vector with...double-stranded DNA transgene is incorporated into a large complex of both viral and cellular proteins and trafficked...titering IDLVs include detecting vector particle components or performing quantitative PCR titration of vector...

  14. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...lacY1 proA2 rpsL20 xyl-5 leu mtl-1 Stbl4   Electrocompetent cells for cloning and storage of unstable ...XL10 Gold Tetracycline and Chloramphenicol High competency cloning and propagation of large plasmids, ligated...tables are by no means exhaustive! For a more comprehensive list and additional information, visit OpenWetWare's...of Bacterial Expression Systems. Check out our companion post describing protein expression strains, where...

  15. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...certain Cas enzymes are less compatible with the bulkiness that accompanies mispairing of bases. To reduce...unnecessary for many applications, it is the only comprehensive method available. Targeted sequencing approaches...must be quantified slightly differently. For a comprehensive approach, RNA-seq would be an appropriate method...

  16. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...helps all viewers, especially if you have some complex patterns with several overlapping targets to interpret...you add the secondary antibody, it should get completely washed away and leave no signal. If you see lots...samples where you label only one target in each. Compare these individually-labeled samples with the results...deep phenotyping and spatial analysis of cells in complex tissues. Proceedings of the National Academy of...

  17. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ...expression, and is flanked (“floxed”) by two sets of incompatible recognition sites (e.g., loxP and lox511) in... Han, 2016). Not so long ago, however, it was completely unclear which subpopulation of DA neurons could... "Two possible roles of bacteriorhodopsin; a comparative study of strains of Halobacterium halobium differing...PMCID: PMC5111696. Towne, Chris, and Kimberly R. Thompson. "Overview on research and clinical applications...

  18. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    ...equipment on a lab bench. The best part? Less complicated math. Bioengineering is a field that has been...achieve whatever goal you’re aiming for. When you complete each cycle, your best-performing mutant is usually...or predicted structure of your antibody-antigen complex as a template. This structural framework then guides... a plate-based format, making it suitable for comparing relative affinities across mutants. Cell surface...

  19. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...could avoid potential off target artifacts by comparing the phenotypes of multiple gRNAs. Targeting different...lanes 0.3 % 3.0, Figure 5, lane 2) is faint, so comparing it to the undigested control (Figure 4, lane 0...instead of agarose gives much sharper bands. I compared two MOIs of gRNA transduction to see if that had...testament to how far genome engineering has come. For comparison, in 2007 it took about 3 months to knockout a...

  20. Addgene’s First AAV Data Hub Challenge

    Type
    Blog Post
    ...your data by Nov. 12 to compete. The top ten participants will be invited to compete in our speed round data.... Bobae An will host a Q&A session after the competition, where you can get all your burning AAV questions...
Showing: 541 - 560 of 950 results