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We narrowed to 1,027 results for: LEA

Showing: 441 - 460 of 1027 results

  1. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...vector via Golden Gate cloning, perform an exonuclease cleanup, transform, plate, and pick. In short, the...found instead ones that were… pretty close, or at least close enough, you’ve found yourself with a common...Fragmid uses Bbsl, a Type II restriction enzyme that cleaves outside of its recognition sequence. This means...in duplicate, they were able to confirm that at least one plasmid was the correct sequence for every individual...

  2. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    ...hatching into their adult phase. It is important to clear out the parent crosses from the container before...sharing-oriented, and there are many resources to help you learn more about fly biology and fly-based experimental... and gene expression. In the next post, you’ll learn the fundamentals about how many of these resources...between pathogens and their hosts. It’s not always clear which side he roots for. You can try to convince...Methods for Site Specific Recombination in Drosophila Learn about Cre-Lox Resources on Addgene.org Find Drosophila...

  3. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...Identifying which genes are important for a phenotype can lead to a wealth of additional experiments investigating...consists of two components: the non-specific endonuclease (Cas9) and a single stranded guide RNA (gRNA...to enrichment or depletion of specific gRNAs can lead to skewed results. Selecting a cell type - Theoretically...validation practices, CRISPR libraries can help you learn a lot about your phenotype of interest. For more...Lentiviral CRISPR Libraries” (McDade et al., 2016). To learn a bit about how CRISPR/Cas9 can be used in other...

  4. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...experience, I was still incredibly nervous about leaving academia. A lot of academics tend to view industry...postdoc experience, I’m going to share what I’ve learned and hopefully make your decision a little easier...publications, which may take a longer time to acquire. Learning about drug discovery: Unless your academic lab...market usually takes over a billion dollars and at least a decade of work. The number of steps, decisions...an academic postdoc, you’re often required to at least attempt to write grants. If you don’t enjoy or excel...

  5. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ... current yellow fluorescent proteins tend to photobleach rapidly, which hinders their use for long-term...compact size and ability to generate heritable edits (learn more in our recent blog post on TnpB). Similar to...TnpB (A) and Cas9 (B) genome editors. Effector nucleases are shown bound to the target DNA, recruited by...tag (ALFA-tag, or At) for easy visualization. A cleavage site for endogenous BACE1 protease allows VAMP2...cell membrane while AF, fused to a payload, is released into the synapse. Once AF binds to the postsynaptic...

  6. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

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    Blog Post
    ...Chromoproteins also enhance corals’ resistance to mass bleaching, the loss of their symbiotic algae, during heat...rhamnose-inducible promoter. The resolution step leads to the desired mutation and excision of the vector...mammalian cell expression here   Now that you’ve learned what chromoproteins are and how they can be used...blog Find our Fluorescent Protein 101 blog series Learn more about FRET Find protocols and tips for making...Blanquet RS, Phelan MA (1987) An unusual blue mesogleal protein from the mangrove jellyfish Cassiopea ...

  7. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...dependencies  Take, for instance, a recent preprint released by the Boehm and Beroukhim labs at the Broad Institute...genetic dependencies of individual tumors, which can lead to new cancer targets and related biomarkers . Data...strongly correlated genetic dependencies that were clearly associated with European or East Asian ancestry... accounted for in the CRISPR guides they used, leading to mismatches between the guides and the lines’...undercounted by approximately 20%. In some cases, this may lead researchers to deprioritize genes that could be ...

  8. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    ...and mascot Blugene (check back on December 5 to learn more about this!). I’ve assembled a series of screenshots...filter, this one from the ‘Requests’ category, will leave us with a single plasmid to consider. If you get...search. Simply click the ‘X’ next to any filter to clear it. Here’s a list of available filters to refine...great improvement for our website and we hope it leads you to exactly what you need with every use. If ...redesign. Additional resources on the Addgene blog Learn more about other features on Addgene's website ...

  9. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...our CRISPR Featured Topic Page Learn about RNA Targeting with Cas13a Learn about CRISPR DNA Base Editing...contain the RuvC and HNH domains responsible for DNA cleavage, so they cannot directly edit the genome. A Cas13...They hoped to find a stably folded ortholog that cleaved RNA robustly, unlike LwaCas13a, which must be stabilized... multiplexed manner to edit multiple RNAs. It’s clear that CRISPR RNA editors will open new doors for ...

  10. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...contributed by guest blogger, Luke Lavis, a Group Leader at the Janelia Research Campus, Howard Hughes Medical... photoactivation, etc. These impressive advances lead to an obvious question: In this age of GFP and its...fluorophore can emit a limited number of photons before bleaching. The amount of information one can extract from...Luke D. Lavis. Luke D. Lavis is currently a Group Leader at the Janelia Research Campus, Howard Hughes Medical...PMC3246539. Additional Resources on the Addgene Blog Learn About Other Protein Tags Catch Up on Your Fluorescent...

  11. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...Resources on the Addgene Blog Learn to do Restriction Cloning Try SLIC cloning Learn about Gibson Assembly ... One key difference is the plasmid DNA must be released from the bacteria in order to serve as PCR template...reaction. You just need to lyse the bacteria to release the plasmid DNA by either briefly boiling the sample...Read about Choosing a Molecular Cloning Technique Learn to Perform a Bacterial Transformation Deposit Your...

  12. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...your cells of interest. Cas9 RNPs are capable of cleaving genomic targets with similar efficiency as compared...levels shortly after transfection, and are quickly cleared from the cell via protein degradation pathways....known as nucleofection, which forms pores in the nuclear membrane, allowing for entry of a DNA template..... "Receptor-Mediated Delivery of CRISPR-Cas9 Endonuclease for Cell-Type-Specific Gene Editing." Journal...the Addgene Blog Subscribe to CRISPR blog posts Learn about other CRISPR delivery techniques Find tips...

  13. DNA Purification Without a Kit

    Type
    Blog Post
    ...cells are lysed, RNAs and proteins are removed to leave you with DNA. This DNA is then washed, and either...purify DNA, cellulose also efficiently binds, or at least entraps nucleic acids. The Botella lab took advantage...around. Finally, purified total nucleic acids are released by dipping the cellulose directly into a PCR buffer...columns, suggesting that columns can be reused at least five times. DNA purification kits don’t come with...used plasmid purification columns could also be cleaned with 1M phosphoric acid and re-used, but trace ...

  14. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...visualizing promoter activity, and much more. Read on to learn more about GFP, how scientists have evolved this...exist with only slight variations separating them.  Please note that many fluorescent proteins found on the...F64L; S65T, Y66W; N146I; M153T; V163A; A206K Cerulean F64L, S65T, Y66W, S72A, Y145A, H148D, N149I, M153T... Y145A and H148D Stabilizes the structure of Cerulean derivatives F99S, M153T, V163A Improves folding...visualize the effects of alterations to this activity. Learn more about optogenetics at OpenOptogenetics and ...

  15. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...different chromatin states. Alexander Stark and colleagues at the Research Institute of Molecular Pathology...Initiative, led by Dominic Esposito, has recently released a collection of 360 Ras pathway plasmids to the...Drosophila cells, targets 13,501 fly genes with at least three independent gRNAs, each chosen strategically...vacuolar) pH environments that inactivate SEP and leave only DsRed to fluoresce (1). With variants that ...effects on gene regulation. Professor Zoldos and colleagues from the Department of the Biology, Division ...26912792 FusX-TALEN assembly system TALEs as nucleases (TALENs) are genome-editing tools that are deployed...auxotrophies or using different auxotrophic backgrounds can lead to disparate physiological effects and complicate...

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

    Type
    Blog Post
    ...replicon of pTargetF may exhibit a more serious leaky expression in BL21(DE3), which we presume caused...that indeed the gRNA-pMB1 had a slightly higher leaky expression in BL21(DE3) that resulted in a lower...a sacB gene renders it beneficial for plasmid clearance. It is more convenient to update the spacer(20nt...Anhydrotetracycline (aTc) induces expression transposases leading to increased integration frequency. (C) Increasing...rounds of transfer on transposition-inducing agar lead to higher cargo copy numbers. (D) The effect of ...

  17. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...integrating neuronal signalling and triggering release of the stress hormone norepinephrine to promote...are chemogenetically activated, norepinephrine release causes a freezing response, a form of immobility... the NAc, where enhanced glutamate transmission leads to locomotion sensitization. (for review see van... as sleep, food intake, mobility/coordination, learning, and memory. We’ll cover these in a  future blog...Guide to Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

  18. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...tips from an AAV rookie  Learn about some considerations when using AAVs Learn more about using AAV as ...complex (LY6A) as a receptor for PHP.B. Read on to learn how the design of PHP.B may have contributed to ...182-188. PubMed PMID: 29175632. If you want to learn more about how PHP.B and PHP.eB were engineered,...PMC5088052. Additional resources on the Addgene blog Learn more about PHP.eB, and PHP.S, a peripheral nervous...

  19. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...remains unclear whether the FP misfolding impacts fusion protein function and this could lead to under...localize [4]. Most investigators are surprised to learn that FPs are often poorly suited for cellular compartments...glycosylation, disulfide bond formation, and proteolytic cleavage. FPs localized to organelles are equally susceptible... dark fluorescent proteins in your experiments, please consider the following: How can one ensure that...

  20. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...bit of a steep learning curve and many opportunities for mistakes. The easiest way to learn optogenetics... is an approximation of reward or the sense of pleasure that the animal perceives through taking a drug...University.  His interests include the role of learning and memory in opioid addiction and science communication...Kheirbek, Mazen A., et al. "Differential control of learning and anxiety along the dorsoventral axis of the...
Showing: 441 - 460 of 1027 results