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We narrowed to 850 results for: met

Showing: 321 - 340 of 850 results

  1. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...your viral vector solution. A variety of titering methods have been developed to determine the titers of ...precision and variability, qPCR is the more widely used method due to the accessibility of instruments in standard... terminal repeat sequences. Human Gene Therapy Methods, 23(1), Article 1. https://doi.org/10.1089/hgtb... of AAV Serotype 1 Vectors. Human Gene Therapy Methods, 28(1), Article 1. https://doi.org/10.1089/hgtb...titers by droplet digital PCR. Human Gene Therapy Methods, 25(2), Article 2. https://doi.org/10.1089/hgtb...

  2. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...Characterization Methods for Quality Control Testing of Recombinant Adeno-Associated Viruses. In: Methods in Molecular... for the contaminant is Addgene plasmid #44362. Method validation, limitations, and future plans How did...another AAV vector with AAV-68544 to validate our method. We then extracted the DNA and processed the mixed... start, but to determine the sensitivity of our method we wanted to see how low we could go. We prepared...

  3. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...   Knockout/Knock-in plasmids Gene targeting methods also make it possible to insert, or knock-in, any...chosen promoter responds. You can also use this method to tag an endogenous protein with GFP, as seen ... given locus.   Conditional knockouts The methods and plasmids described in the preceding sections...The future of knockout plasmids Although these methods have been used to create many knockout cell lines...Addgene depositors have developed new CRISPR-based methods for various knock-ins, discussed in our CRISPR ...

  4. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...option, but the equipment needed for EM is not something you find in the average cell and molecular biology... continuous lines in pre-expanded samples can sometimes take on a fragmented appearance post-expansion...ultrastructures using expansion microscopy (U-ExM). Nat Methods 16:71–74. https://doi.org/10.1038/s41592-018-0238...fluorescent Fab fragment secondary antibodies. Methodsx 9:101796. https://doi.org/10.1016/j.mex.2022.101796...principles and uses in biological research. Nat Methods 16:33–41. https://doi.org/10.1038/s41592-018-0219...

  5. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

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    Blog Post
    ...sgRNA-level metrics to assess library performance in viability screens. Based on these metrics the Brunello... CRISPRi libraries with 10 sgRNAs in the sgRNA metrics, confirming again that sgRNA design plays a significant...dCas9 and its RNA directed binding site. These methods include the fusion of an activation domain such...of multiple copies of VP16 termed the “Sun Tag” method (Tanenbaum et al., 2014). The sgRNA or tracrRNA...

  6. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ... for genetic studies (e.g. using mariner), as a method of tagmentation for next-gen sequencing pipelines... insertions are not currently possible, unlike methods involving homologous recombination. Additionally...Additionally, these transposon also exhibit something called target immunity, whereby multiple proximal insertions..., 2020), to multi-copy genomic integration of metabolic cassettes (Zhang et al. 2020); however, the technology...of our CRISPR blog posts Check out other CRISPR methods for bacteria Resources on Addgene.org Visit the...

  7. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...in large enough quantities (and the correct stoichiometric ratios) to generate useable data. What’s more...easier said than done. Fortunately, a complementary method exists for visualizing protein-protein interactions...? Bimolecular fluorescence complementation is a method for detecting protein-protein interactions that..., 2006). One of the first BiFC studies used the method to interrogate the subcellular localization of ... other BiFC is a convenient, fluorescence-based method for directly visualizing PPIs in cells. The ease...

  8. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...Tips below), and by validating leads with other methods. More common are false negatives; up to 75% of ...possibility for problems with one or more of these parameters can lead to a high number of false negatives ...five independent protein interaction detection methods (6). In order to bind DNA and activate a reporter...gene expression in Escherichia coli and yeast. Methods Enzymol. 1983;100:293-308. PubMed PMID: 6312261...Benchmarking of the yeast two-hybrid system. Nature Methods. 2010; 7:667-668. Pubmed PMID: 20805792. 7. Stynen...

  9. Plasmids 101: Plasmid Incompatibility

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    Blog Post
    ...It is also worth noting that plasmids can be symmetrically incompatible, meaning that both plasmids are... from the cell with the same probability, or asymmetrical, where one plasmid lost from the cell at a higher... higher rate than the coresident plasmid. Asymmetric plasmid loss occurs for several reasons, including...high copy incompatible plasmids that lead to asymmetrical plasmid loss of the usually large, low copy ...Suzuki K, Yoshida K (2002) A novel plasmid curing method using incompatibility of plant pathogenic Ti plasmids...

  10. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...coli B and W strains. The optimized plasmid curing method was much simpler and took only ~32 hours instead...serve to interrupt genes (or gene clusters) in metabolic engineering. So far, we’ve used the MUCICAT system...when compared to the industrial strain. In our metabolic engineering case study of N-acetylglucosamine ...Multiple induction systems to choose from. Ready for metabolic engineering with improved cargo dedicated to gene...from our CRISPR topic overview Find other CRISPR methods for bacteria Find all articles about microbiology...

  11. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...What is Rett Syndrome? Caused by a mutation in the methyl-CpG binding protein 2 (MECP2) gene, Rett Syndrome...et al., 1999). We now know that MECP2 binds to methylated CpG dinucleotides and CAC trinucleotides during...caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet 23:185–188 . https.../13810 Amir RE, Zoghbi HY (2000) Rett syndrome: Methyl-CpG-binding protein 2 mutations and phenotype-genotype...after Intracisternal Delivery. Molecular Therapy - Methods & Clinical Development 5:106–115 . https://doi....

  12. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...easier, faster, and more efficient than previous methods. Still, generating reliable results from CRISPR...sequences, and ICE will do the rest. There are no parameters that need optimizing and no complicated steps...yellow check mark indicates that a particular parameter was automatically adjusted to generate ICE results...Successfully analyzed samples will display the following parameters: Sample - The unique label name that you provided...gene. The analysis can be sorted by any of the parameters displayed in the summary table. You can also ...

  13. CRISPR 101: Homology Directed Repair

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    Blog Post
    ...non-conservative and conservative. The conservative methods are characterized by use of a homologous donor ...break-induced repair (BIR). The non-conservative method is single strand annealing (SSA), an error prone...Alterations. Edited by Doudna J and Sontheimer EJ. Methods in Enzymology Vol. 546; (2015). PubMed. Optimization...catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Richardson CD, et al. Nat Biotech....Genome Engineering. 2017.  Easi-CRISPR: a robust method for one-step generation of mice carrying conditional...

  14. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    ...integrins. Six of the 11 IPI antibodies contain RGD-mimetic sequences that imitate this motif and compete with... unconjugated and recommended for use in flow cytometry. Based on the established applications of the ...resulting in cells that elongated and morphed asymmetrically (Jo et al., 2022). By contrast, LVDP-binding...conferred less force, leading cells to spread into symmetrical circles. While elucidating the role of tension...Addgene's Antibody Guide Antibodies 101: Flow Cytometry Antibodies 101: Introduction to Antibodies...

  15. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...minutes, they are perfect for molecular cloning and metabolic engineering.  The E. coli genome has an extensive...address many areas of microbial research like metabolism. Transport, biofilms, and much more. Because ..., breakthroughs in gene modification tools and methods such as CRISPR have vastly improved scientists’...variation between different plant species and metabolic properties affecting adaptations.  Arabidopsis...doi.org/10.1159/000490898 Gibbs, R., Weinstock, G., Metzker, M. et al. Genome sequence of the Brown Norway ...

  16. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ... populations of synapses and deep-brain fiber photometry.  Figure 1: Characterization of iGluSnFR4...higher efficiency and lower indel formation than methods relying on homology-directed repair (HDR), and ...immunoprecipitation (IP), and tandem mass tag (TMT) mass spectrometry (MS) in HCT116 cells (colorectal cancer). The...isolated and identified with tools like mass spectrometry, providing a snapshot of interacting proteins...Bartom, E. T., & Shilatifard, A. (2024). TurboCas: A method for locus-specific labeling of genomic regions ...

  17. A Control for All Seasons

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    Blog Post
    ...is out of your hands, it’s helpful to focus on something controllable, like experiments (and their controls...use a preadsorption control to confirm. In this method, the primary antibody is incubated with its antigen...that the target protein was measured on.    Flow cytometry In addition to the controls listed above, viability...compensation controls are recommended for flow cytometry. A viability control allows you to detect dead...fluorophore and are critical to analyze multiplex flow cytometry experiments. We hope that this blog post will ...

  18. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ... using CRISPR is easier than more conventional methods. Time and money savings come from the fact that...mouse editing when compared to more conventional methods. First, one can work with almost any strains of... be $50,000. Finally, compared to conventional methods of genome engineering, CRISPR is much more efficient...2016) GONAD: A Novel CRISPR/Cas9 Genome Editing Method that Does Not Require Ex Vivo Handling of Embryos...101: Mammalian Expression Systems and Delivery Methods Learn about Mouse Work on Exercise and Depression...

  19. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...create stable transgenic cell lines. The traditional method of making stable cell lines is to transfect, select...piggyBac™ gene editing system compare with other methods of gene editing? There are primarily two other .../loxP) to remove the donor cassette. While this method can be effective, a loxP site is left behind in...Pluripotent Stem Cells with Seamless Gene Editing.,” Methods Mol. Biol., Apr. 2016. PubMed PMID: 27075976. 4.... J. Adams, “A high-throughput splinkerette-PCR method for the isolation and sequencing of retroviral ...

  20. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ... you can help ensure that you understand the methodology and can have confidence in the reagents and outcomes...is preferable to finding out you've been doing something all wrong or that a reagent has not been convincingly...feedback from experts. If someone crosses out something, find out why. Ask successful writers about their...exercise. I highly recommend running or biking or something that demands your attention. Eat well- a diet... help maintain your health. Have a hobby- do something that gets you away from thinking about the lab...
Showing: 321 - 340 of 850 results