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We narrowed to 795 results for: Kin

Showing: 441 - 480 of 795 results

  1. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...in brackets and add the upstream and downstream flanking sequences to their input string (Figure 2). This...prediction models. In addition, we provide options for nicking gRNAs (used for PE3 and PE5 prime editing strategies...strategies) based on a DeepSpCas9 score ranking (Kim et al., 2019). PRIDICT saves time and resources by ...

  2. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ... localization, cell structure, intracellular trafficking, and protein turnover rates. Additionally, by...fission/fusion, nuclear import, and neuronal trafficking. Although FPs have enabled discovery of many ...cellular mechanisms, there are some limitations to working with FPs. Overexpression of fluorescently tagged...

  3. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ... I’m not going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs...the viral particles of the AAV serotype you’re working with are primarily found in the culture media, .... If you’re new to creating gradients, practice making the iodixanol layers before doing your first purification...

  4. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...components of the indicator, which can make the kinetics of voltage sensing to signal output slightly slower...fluorescent signaler. The pro of this system is the kinetic response time from signal input to readable output...FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component counterpart and...

  5. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...the FP (fused to the protein of interest), thus tracking the localization of that biomolecule inside living...eel was shown to mature independently of oxygen, making suitble for use in anaerobic conditions. Maturation...fluorescent protein should I use?" And if you're looking for more fluorescence microscopy tools and techniques...

  6. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...domain proteins, they can be expressed in bacteria, making nanobodies a recombinant and renewable anti-IgG...available nanobodies from Pleiner et al. Before picking a secondary nanobody, it’s important to review ...expression plasmids here Additional Resources Looking for monoclonal primary antibodies to use with the...

  7. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...does not do). Furthermore, molecular biologists working with vectors are, in most cases, interested in ... the viral integrase protein (IN) during step 3 (King et al., 2008) (Hu & Temin, 1990). At step 3, the....1016/0022-2836(73)90277-5. PubMed PMID: 4355596 King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C....

  9. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    Published
    Jan. 16, 2020, 2:18 p.m.
    ...our samples 1:6 million to 1:25 million.  After making the dilutions, they are transferred to a new plate...performing ddPCR Clean everything You must have a clean working area to titer your AAV by ddPCR. Droplet digital...generator, you should avoid pipetting too quickly when making your virus dilutions. This will reduce aerosols...

  10. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...dye, mauvine, was discovered in 1856 by William Perkin. His discovery set off a flurry of activity and...labeling strategies and improved fluorophores are making chemical dyes increasingly attractive and accessible...Medical Institute. He would rather be in the lab making new fluorophores for advanced imaging modalities...

  11. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

    Type
    Blog Post
    Published
    Sept. 9, 2025, 1:15 p.m.
    ...enhancer candidates. To use these sequences for making functional genetic tools, enhancers were nominated...Institute are committed to just that, and their painstaking efforts towards screening thousands of enhancer...Hooper, M., Omstead, V., Vargas, S., Lerma, M. N., Taskin, N., Weed, N., Laird, W. D., Bishaw, Y. M., Bendrick...

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

    Type
    Blog Post
    Published
    July 30, 2024, 1:15 p.m.
    ...reliable and robust high quality viral vector service, making an array of ready-to-use viral vectors available.... I would be remiss to talk about them without looking at what they’re delivering and where they’re delivering...of our most popular ready-to-use viral vectors, making up almost 10% of our requests by 2019 (Figure 5A...

  13. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ... replication and regulation in bacteria Before looking at an example plasmid, let's review how E. coli...different ways: methylating newly synthesized DNA, blocking binding sites, or trading out ADP for ATP to re-activate...mRNA and prevent this pseudoknot from folding, blocking replication. Created with BioRender.com. Plasmids...

  14. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    Published
    May 21, 2024, 1:30 p.m.
    ...information to explain why you are interested in working for that organization or group. This is particularly... of what to put in your cover letter, start by looking at the requirements in the job description. For... good place to state why you are interested in working for this lab and/or company. Remember to proofread...

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

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...through smart engineering Different FLP versions – making FLP efficient The first version of FLP discovered...proteins needed for later experiments (cellular tracking for instance) can be controlled with FLP-FRT. ... Cardin, Jessica A. et al. 2009. “Driving Fast-Spiking Cells Induces Gamma Rhythm and Controls Sensory...

  16. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...when 2 mismatches are present. The protospacer flanking sequence (PFS) for LshCas13a, which is analogous...have utilized dCas13 fusion proteins for imaging, tracking, modulating splicing, and regulating expression... in length and does not require a specific PFS, making it a very flexible cleavage system. LwaCas13a is...

  17. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    Published
    April 11, 2017, 2:30 p.m.
    ... of how they actually “report” as biosensors. Making aptamers glow The flexibility of aptamers means...fluorogen and increase its fluorescence, essentially mimicking GFP.  Though the aptamer, called Spinach, required... Eric J. Perkins...

  18. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ...BP Reaction takes place between the attB sites flanking the insert and the attP sites of the donor vector...gene! STEP 2: Generate an Expression Clone When making the expression clone, it is important to choose...fragments, cDNA or Genomic DNA and is available for all kind of organisms from mammals to E. coli. Glossary of...

  19. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    Published
    Aug. 13, 2019, 12:58 p.m.
    ...zebrafish embryo develops outside of the mother, making them ideal for studying vertebrate embryogenesis... target mRNA inhibit its translation either by blocking initiation or splicing. (RNAi, in contrast, engages...unprecedented views into cognitive processes like decision making (Kim et al, 2017). “Brainbow” fish lines have been...

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

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ...speech, coordination, purposeful use of hands, and walking. They may exhibit slow head growth, develop seizures...medical journal. Decades later and after years of painstaking work trying to isolate the causative gene for...:2<147::aid-ajmg6>3.0.co;2-o Baker SA, Chen L, Wilkins AD, Yu P, Lichtarge O, Zoghbi HY (2013) An AT-Hook...

  21. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ... delivery to only a few cell types.  If you’re looking for more control in the cell types your lentiviral...retrograde direction from the synapse to the cell body, making it a great tool to study neuronal connectivity....HY, Tortorici MA, Veesler D, Murphy M, Pettie D, King NP, Balazs AB, Bloom JD (2020) Protocol and Reagents...

  22. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    Published
    Nov. 10, 2016, 3:30 p.m.
    ... month without a new, drug-resistant ‘superbug’ making headlines in one part of the world or another. ...that only infect bacterial cells), or a bacterium picking up resistance genes from it’s environment (from...resistant bacteria to over-take the population and making them more difficult to treat in the future. Patients...

  23. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...a bit of tuning before you know your system is working as expected. Finally, for each gene you want to... scorer 1.0 (Now 2.0) (Chari et al., 2015) and picking whichever guides are closest to the transcription...Ansari R, Harper S, Jackson DA, McRae R, Pooley R, Wilkinson P, van der Meer D, Dow D, Buser-Doepner C, Bertotti...

  24. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...that particular region. No matter where you are looking, you can either scan the genome yourself for potential...leading to degradation that can set you back days. Working with plasmid DNA is much easier and doesn’t require...slightly different method to genotype their pups. Taking a toe, a finger, a tail tip, or a combination of...

  25. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    Published
    Oct. 1, 2015, 2:30 p.m.
    ...we are interested in circuits that mediate drug taking behavior. If a circuit reinforces behavior (activation...sense of pleasure that the animal perceives through taking a drug. The ideal behaviors to test reinforcement...amygdala to nucleus accumbens facilitates reward seeking." Nature 475.7356 (2011): 377-380. Pubmed PMID:...

  26. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    Published
    June 17, 2025, 1:15 p.m.
    ...sequence (Figure 2A). Since you don’t need additional flanking sequences, shRNA plasmids are a bit smaller, with... than miRNA plasmids, which require additional flanking regions comprised of upstream and downstream genomic...between the red boxes. Highlighted blue regions show flanking genomic sequence. Created on BioRender.com. ...

  27. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    Published
    April 4, 2024, 1:15 p.m.
    ...cells of interest. In many experiments you’ll be working with a heterogeneous cell population, for example...While flow cytometry can be used for virtually all kinds of experiments and research fields, for any experiment...investigating negative regulators of T cells and cytokine signaling in tumor immunity. When he's not in ...

  28. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    Published
    March 5, 2019, 1:08 p.m.
    ... this, similar systems have been developed for tracking various cell types and organisms and for improving...recombinase. The XFPs in Flybow are membrane-tethered, making this system useful for visualization of cellular...construct was then used to develop a system for tracking the fate of Langerhans cells. In the Ubow mouse...

  29. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...dependency on magnesium and higher thermal stability, making it a potentially more useful tool for in vivo studies...much potential for growth as RNA biology, we are looking forward to reports on the next additions to aptamer...bright, high-affinity complex for RNA labeling and tracking." ACS chemical biology 9.10 (2014): 2412-2420....

  30. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...false positives observed in the cloning process, making it easier to produce the constructs you need. Expression...to simplify the protein purification process by making it easy for a researcher to quickly generate N-...carefully chosen to perform statistically relevant benchmarking of cell-free expression systems, and tested ...

  31. Evolution of Lab Techniques

    Type
    Blog Post
    Published
    June 21, 2016, 2:30 p.m.
    ...nucleotides, called dideoxynucleotides (ddNTPs), lacking the 3’ hydroxyl group required for the extension...sequencing reactions and may use Sanger sequencing. Looking to the future Southern blotting, restriction mapping...mouse lines, such as BAC transgenic or traditional knockins/knockouts, a thing of the past. In the sequencing...

  32. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ... however NLuc’s emission peak is at 460 nm thus making it non-optimal for in vivo imaging. The solution...about to create an in vivo imaging reporter by linking two CyOFP1 molecules to NLuc (CyOFP1-NLuc-CyOFP1...the fluorescent donor for a bioluminescent donor, making BRET probes. Here the bioluminescent donor is unaffected...

  33. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ...adds a repressible element to this binary system (making this a repressible binary system). The GAL80 repressor...PMID: 1319065; PMCID: PMC49329. Triezenberg SJ, Kingsbury RC, McKnight SL. Functional dissection of VP16... Resources on the Addgene Blog Quick guide to working with Drosophila Terminators and PolyA Signals FLEx...

  34. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ...in common? The three most important items when looking for a house and choosing an AAV are… location, .... So how do you choose the right serotype? Keep asking questions!  Table 1: AAV Serotypes and Tropism ...choice of AAV will depend upon which section you’re looking at (striatum, hippocampus and cortex) within the...

  35. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    Published
    Dec. 15, 2016, 3:57 p.m.
    ...interest and 50 nucleotides of homology to regions flanking the target site).  The best applications for dsDNA...Daiguan Yu, Hilary M. Ellis, E-Chiang Lee, Nancy A. Jenkins, Neal G. Copeland, and Donald L. Court. An efficient...Read about Homology Directed Repair Use Knockout/Knockin Plasmids to Disrupt Genes in Mammalian Cells CRISPR...

  36. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    Published
    March 6, 2018, 1:49 p.m.
    ... a method relies on accurate ligation of nicks lacking gaps or mismatched base pairs (as in the case for....6 (2004): 757-764. PubMed PMID: 15582400. 2. Tomkinson, Alan E., et al. "DNA ligases: structure, reaction...: 16464020.  3. Ellenberger, Tom, and Alan E. Tomkinson. "Eukaryotic DNA ligases: structural and functional...

  37. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...RNase III enzyme named Dicer (Figure 1). The plant kingdom has expanded its repertoire of Dicer enzymes and...silencing effect through targeted mRNA degradation, blocking translation, or chromatin modifications of complementary...researchers studying plants and other organisms lacking this innate immunity need not worry and have benefited...

  38. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    Published
    Feb. 25, 2020, 2:15 p.m.
    ... and SciFi.  Scientists around the world began looking for the magic molecule that serves as the code ...insatiable appetite of the moviegoers for dinosaurs wreaking havoc in a theme park.  Released in 1997, GATTACA... at University of California San Diego. She is working in collaboration with Prof. Alexis Komor and together...

  39. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    Published
    May 28, 2024, 1:45 p.m.
    ...the better the detection of the target marker. Looking at Table 1, we can recognize a couple of fluorophores...brightness levels are often not that great. When thinking of terms like "very bright" vs. "dim" you might...investigating negative regulators of T cells and cytokine signaling in tumor immunity. When he's not in ...

  40. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...the template strand. Therefore, I suggest that picking a crRNA complementary to the template strand could....For ssODN design, remember to incorporate PAM-blocking mutations (e.g. convert the NGG PAM to NGC in ...PCR genotyping (Sanger sequencing); use primers flanking the tag sequence and expect a size-shift in the...
Showing: 441 - 480 of 795 results