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  1. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    ...the 3rd-generation packaging systems offer the maximal biosafety available for the technology. The minor... divided as follows: Transfer vectors contain minimal cis-acting HIV components: LTRs, PPT, RRE, and ...vectors are SIN. Packaging plasmid(s) contain the minimal number of HIV genes required for virus production...
  2. Antibodies 101: The Four ELISAs and When to Use Them

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    Blog Post
    ...adapted it for a host of different proteins and small molecules from a variety of sample types (Engvall... competitive or inhibition ELISA when measuring small molecules that cannot bind efficiently to two antibodies... develop assay Competitive Appropriate for small moleculesReduced chance of cross-reactivity Less...
  3. Hot Plasmids Spring 2024

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    Blog Post
    ...impacting prime editing and found the protein La (a small RNA-binding exonuclease protection factor) promotes...2024) Improving prime editing with an endogenous small RNA-binding protein. Nature. 2024 Apr 3. doi: https...trans-splicing. By splitting a large gene into smaller fragments, delivering them by individual AAVs, ...
  4. Viral Vectors 101: Preparing Pooled Libraries

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    Blog Post
    ...surprised at how this powerful tool is in such a small package. But don’t let the simple presentation deceive...freeze-thaw cycles needed when using the virus. A small aliquot is used to determine functional titer with...aliquot from an amplification done by Addgene.) This small aliquot is not enough to transfect producer cells...
  5. Plasmids 101: Gateway Cloning

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    Blog Post
    ...lentiviral expression of the human KRAS gene in mammalian cells. STEP 1: Generate an Entry Clone There are...expression level, and experimental purpose. For mammalian lentiviral expression, we could use a vector like...and is available for all kind of organisms from mammals to E. coli. Glossary of Gateway cloning vectors...
  6. Plasmids 101: Plasmid Incompatibility

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    Blog Post
    ...where one plasmid outcompetes the other, usually a smaller, higher copy plasmid can more effectively titrate...Environmental Plasmids blog). Scientists have designed small, high copy incompatible plasmids that lead to asymmetrical...2012) Bacterial plasmid partition machinery: a minimalist approach to survival. Current Opinion in Structural...
  7. An Inside Look into Addgene's Dev Team

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    Blog Post
    ...blind spots. Blind spots often result in less than optimal user experience and the overall soundness of the...development. Scrum teams plan and break up work into small, well defined chunks, called stories, and work on... to the story being scrapped or broken up into smaller stories. Since this occurs before a single line...
  8. CRISPR-mediated Plant Base Editors

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    Blog Post
    ...studies demonstrated successful base editing in mammals, but there is a lack of base editors for plant ...PAM). ABE7.10 can edit target 'A' if it lies in a small genomic window: at protospacer position 4-8 counting...Agrobacterium-mediated transformation. We also constructed smaller vectors suitable for transient assays using protoplast...
  9. Plasmids 101: Codon usage bias

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    Blog Post
    ... Pechmann and Frydman found that tracts of non-optimal codons are associated with specific secondary structures...coli strains express a variety of tRNAs that are normally found at low abundance in E. coli. The advantage... Frydman. "Evolutionary conservation of codon optimality reveals hidden signatures of cotranslational ...
  10. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...optimum of 30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular...populations) within neuronal networks in living animals. To accomplish such cell specific manipulations.... “Conditional Site-Specific Recombination in Mammalian Cells Using a Ligand-Dependent Chimeric Cre Recombinase...
  11. CRISPR 101: Targeting RNA with Cas13a (C2c2)

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    Blog Post
    ...COVID-19 (Joung, Ladha et al., 2020). Cas13a in mammalian cells The Zhang lab made the exciting discovery...discovery that some Cas13a orthologs can function in mammalian and plant cells (Abudayyeh et al., 2017). They...superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can mediate cleavage with...
  12. 5 Reasons to Use Reddit for Science Communication

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    Blog Post
    ...for permanent reference by the Winnower. The only small downside to the AMA from my end as an organizer/...of great stuff there anyway), but this is just a small quibble. If you are not too worried about selling...resources to the basic questions and to provide a small bit of support (you are not alone!) for those going...
  13. Plasmids 101: Control Plasmids

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    Blog Post
    ...reagents and process. It is important to note that optimal experimental conditions, including how much plasmid...backbone as Plasmid A—that does not target any mammalian gene. This non-targeting control is critical to...a gene, you should design a control that shows maximal activation. Likewise if you are trying to repress...
  14. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    ... refer to antibodies and 'staining' to refer to small molecule dyes. Indirect labeling While antibodies...of those CD8+ T cells, require the cells to be minimally altered, so you would not be able to use antibodies... Lastly, stains, based not on antibodies but on small molecule dyes, are useful, most notably when it ...
  15. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...The new SuperPLDs work with high efficiency in mammalian cells and in vitro, opening up a wide realm of...directed-evolution methods, the activity-based selection in mammalian cells used here could be an attractive strategy...Activity-based directed evolution of a membrane editor in mammalian cells. Nature Chemistry, 15(7), 7. https://doi.org...
  16. Deep Mutational Scanning with One Pot Saturation Mutagenesis

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    Blog Post
    ...pool down to plasmids expressing variants with optimal function. High-throughput DNA sequencing is then...ubiquitination activity assays, toxin activity, small molecule binding, growth rate, and many more. See..., Gibson assembly is better suited for creating small, focused scanning libraries. PFunkel: Of prior...
  17. Antibodies 101: Introduction to Immunofluorescence

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    Blog Post
    ...Good fixation maintains sample morphology with minimal impact to the target epitope. There are multiple...Sample Preparation This step can involve multiple smaller steps that all help to ensure that antibodies can...Sample Preparation, this step can involve multiple smaller steps, but the aim is to preserve sample integrity...
  18. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ...deliver the best possible videos for free. We’re a small operation, so sometimes it can be a big challenge...premium, so I had to figure out a solution for that. Normally, shots require me to get in close proximity to...good once. Even when things return to relative normalcy, I’ll be incorporating that mobile monitor stand...
  19. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ...Approach: Viral RNA is extracted and subject to isothermal amplification by primers that target the E and...RNA is extracted from a sample and subject to isothermal recombinase polymerase amplification (RPA) using...contain reverse transcriptase to produce cDNA. Isothermal amplification is done at one constant temperature...
  20. How to Negotiate a Successful Lab Start Up Budget

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    Blog Post
    ...science, and can include: equipment, reagents, animals, glassware, and even people. When developing a ...budget bottom line costs may be rather high. That’s normal, because this is where you can start to identify... modeling and collaborations rather than using animals, then that’s a cost that you can eliminate from...
Showing: 421 - 440 of 812 results