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Showing: 401 - 420 of 844 results
  1. Viral Production at Addgene

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    Blog Post
    ...preparation by precipitation in 10% polyethylene glycol (PEG) followed by centrifugation. Precipitated pellets...i.e., genome-containing) AAV particles within representative vector preparations was quantified with electron...for viral production are propagated in the endA-mutated NEB Stable strain of E. coli. In addition, plasmids...
  2. Hot Plasmids: Fall 2024

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    Blog Post
    ...TBD domains, part of the HB domain, and several mutations to improve solubility and stability (Figure 2)...through eight stages of embryonic development. Representative images of single focal planes through the middle...in natural DNA repair pathways and spontaneous mutations.  Here, the researchers were able to target multiple...abscessus either simultaneously or stepwise, achieving mutation efficiencies of up to about 9% of colonies (102...
  3. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

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    Blog Post
    ... same (forward) or in the opposite (reverse) orientation? The answers to questions like these will determine...sequences, called modules or parts. Assembly is dictated by four-base overhangs that are paired with different... the sequence, normally through the synonymous mutation of a codon. You can domesticate the sites yourself...colony PCR electrophoresis), is free of undesired mutations, and that you have the correct four-base overhangs...
  4. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...determine the presence and quantity of DNA, RNA or a mutation of interest. Together SHERLOCK and DETECTR demonstrate... specificity.  SHERLOCKv2 SHERLOCK had a few limitations when it was first introduced in 2017 so the lab... pre-amplification step allowing for greater quantitation without compromising sensitivity. To detect...SHERLOCK could also detect low-frequency cancer mutations from cell-free DNA fragments as well as health-related...
  5. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ... constant variables that enable the correct interpretation of the effect of the independent variable you...controlled for in order to eliminate any alternative interpretation of the results. Typically, plasmid-based experiments...non-targeting control is critical to the correct interpretation of the results because it provides an important...etc, but outside constraints may impose some limitations and you should be cognizant of these when interpreting...
  6. Plasmids 101: The Promoter Region – Let's Go!

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    Blog Post
    ...sense OR antisense transcript depending on its orientation to your gene. T7lac High levels of gene expression...sense OR antisense transcript depending on its orientation to your gene. araBAD General expression Promoter... promoter with somewhat weak expression. lacIq mutation increases expression of the repressor 10x, thus...sense OR antisense transcript depending on its orientation to your gene Although this list is a great...
  7. Hot Plasmids: Summer 2024

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    Blog Post
    ...glutaraldehyde crosslinking: representative micrograph (C), particle orientation distribution and Fourier-...natively isolated complexes. To address these limitations, the Funabiki Lab developed a technique called...of LEAs may bias the distribution of particle orientations, this could be overcome with methods such as...
  8. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ...post was contributed by Shravanti Suresh from Iowa State University. Since its appearance, SARS-CoV-2 has... fluorescence can be observed under either no excitation light, UV light or LED blue light.  Highlights...to our guest bloggers Shravanti Surest from Iowa State University. Shravanti Suresh is a PhD candidate ...CRISPR/Cas immune systems in the Sashital Lab, Iowa State University. She is actively interested in science...
  9. Viral Vectors 101: Systemic Capsids

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    Blog Post
    ...recombination target sequences, denoted by triangles] facilitate tissue or cell type specificity. An on/off genetic...administered intravenously. To address these limitations, labs have developed techniques like CREATE (...broad):  enhanced coverage for inhibitory versus excitatory neurons Mice, rats Yes (LY6A) Chan et al...AAV.CAP-B10 CNS: balanced coverage of inhibitory and excitatory neurons Mice, marmosets Unknown Brown et... its tropism is relatively more biased toward excitatory neurons than AAV-PHP.eB (Fig. 4c, Jang et al....administration of novel engineered AAV capsids facilitates enhanced transgene expression in the macaque ...
  10. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

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    Blog Post
    ...first generate your desired mutation in mouse ES cells, select for the mutation and inject the ES cells into... models. For simple knockout alleles or point mutations, the 8-10 month process of conventional gene targeting...PAM; if not, you can add a silent PAM or guide mutation. Designing an Easi-CRISPR conditional allele The...
  11. Plasmids 101: Repressible Promoters

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    Blog Post
    ...which can be switched from a default OFF to an ON state, and how you might use these in your research. Today...which can be switched from a default ON to an OFF state, as well as repressible binary systems. In recent...domain VP16. cTA binds to promoters containing putative operator sequences (CuO) (linked in groups of ...QUAS promoter is less leaky than UAS in the basal state, and co-expression of the inducer QF can increase...
  12. Hot Plasmids: Spring 2025

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    Blog Post
    ...hearing about researchers' favorite features and limitations of existing backbones, we're excited to share...5: TurboCas protein labeling compared to immunoprecipitation. The top panel depicts the identification...gene promoter using TurboCas, streptavidin immunoprecipitation (IP), and tandem mass tag (TMT) mass spectrometry...interneurons — those marked by parvalbumin (PV), somatostatin (SST), vasoactive intestinal peptide (VIP), ...
  13. When Fidelity Matters: A frank discussion about ligase fidelity

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    Blog Post
    ...nucleotide sequences with high specificity and quantitative accuracy and even include profiling single nucleotide...Electrochemiluminescent assay for detection of extremely rare mutations based on ligase reaction and bead enrichment."...direct identification of allelic composition of mutated DNA in a mixed population of excess wild‐type DNA...probe for the detection of low-abundance point mutations." Analytical chemistry 86.1 (2013): 900-906. PubMed...
  14. Antibodies 101: Flow Cytometry

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    Blog Post
    ...of the population as a whole, scientists gain statistical power in their observations. However, while flow...Techniques is Best for Me?  An overview of the instrumentation A flow cytometer combines three systems to ...fusions by binding to the protein in its native state.  Antibodies-based flow cytometry can use both direct...cytometry: retrospective, fundamentals and recent instrumentation. Cytotechnology 64:109–130 . https://doi.org...
  15. Your Top Requested Plasmid in 2016!

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    Blog Post
    ... V2.0 from it's predecessor, which had a point mutation in the puromycin resistance gene that made it ...less effective in certain cell lines. This point mutation has been corrected in V2.0. pSpCas9(BB)-2A-Puro...than creating double stand breaks and may better facilitate homology directed repair. You can find all of...
  16. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

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    Blog Post
    ...is limited by definition, the kinome is a well-annotated collection of genes that can provide a useful ...useful for imaging or gene expression assays necessitating the screening of individual plasmids separately... gRNAs that score as hits in pooled screens, facilitating more rapid follow-up. We hope the biological...
  17. Top 10 Open Science Developments of 2013

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    Blog Post
    ...Also in June, the G8 Science Ministers issued a statement after a meeting in London on the global science...available to others got a nine percent bump in citations for their trouble, even when all kinds of other... as much about open access as it does about the state of peer review. 8)     That same issue of Science...
  18. Six Spooky Science Stories and Halloween at Addgene

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    Blog Post
    ...eyestalk and transform them into a pulsating display imitating a moving caterpillar. The worm controls the snail...discovered in Drosophila melanogaster (Gilbert, 2004). Mutations in these genes are lethal, associated with defective..."Tongue replacement in a marine fish (Lutjanus guttatus) by a parasitic isopod (Crustacea: Isopoda)." ...
  19. Supporting Rare Disease Research with BeHeard 2017

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    Blog Post
    ...but many throughout the world are affected by devastating diseases that affect far fewer people and therefore...provide hope to patients suffering from these devastating afflictions. Elia Di Schiavi - Spinal muscular...Nabavi is a Postdoctoral Fellow at the Vancouver Prostate Center and is affiliated with the University of...
Showing: 401 - 420 of 844 results