We narrowed to 786 results for: KIN
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TypeBlog PostPublishedJune 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...
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Tips for Writing a Good Cover Letter
TypeBlog PostPublishedMay 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... -
Viral Vectors 101: Pseudotyping
TypeBlog PostPublishedApril 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... -
CRISPR 101: Targeting RNA with Cas13a (C2c2)
TypeBlog PostPublishedSept. 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... -
Fluorescent Proteins 101: Aptamer Fluorophores
TypeBlog PostPublishedApril 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... -
Plasmids 101: Gateway Cloning
TypeBlog PostPublishedJan. 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... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog PostPublishedNov. 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... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog PostPublishedOct. 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... -
CRISPR Activation: A Practical Guide
TypeBlog PostPublishedAug. 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... -
Tips for CRISPR Gene Editing in Mice
TypeBlog PostPublishedJune 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... -
Important Considerations in Optogenetics Behavioral Experiments
TypeBlog PostPublishedOct. 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:... -
Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior
TypeBlog PostPublishedMarch 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... -
The Fluorescent Vegetables in Aptamer Soup
TypeBlog PostPublishedJan. 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.... -
Plasmids 101: Expressing shRNAs for RNA Interference
TypeBlog PostPublishedJune 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. ... -
Antibodies 101: Introduction to Gating in Flow Cytometry
TypeBlog PostPublishedApril 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 ... -
Multiple Plasmids at a Low Price: Kits from Addgene
TypeBlog PostPublishedSept. 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 ... -
Evolution of Lab Techniques
TypeBlog PostPublishedJune 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... -
Viral Vectors 101: AAV Serotypes and Tissue Tropism
TypeBlog PostPublishedJan. 16, 2025, 3:42 p.m....2008). It can also cross the blood-brain barrier, making it useful for applications targeting the CNS (Issa... Tabebordbar, M., Lagerborg, K. A., Stanton, A., King, E. M., Ye, S., Tellez, L., Krunnfusz, A., Tavakoli...Bertera, S., Zhang, J., Chen, C., Papworth, G., Watkins, S., Trucco, M., Robbins, P. D., Li, J., & Xiao... -
Technologies Enabled by NanoLuc® Luciferase
TypeBlog PostPublishedFeb. 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... -
Plasmids 101: Repressible Promoters
TypeBlog PostPublishedDec. 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... -
RNA Interference in Plant Biology: New Tools for an Old Favorite
TypeBlog PostPublishedOct. 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... -
A History of Genome Engineering in Popular Culture
TypeBlog PostPublishedFeb. 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... -
When Fidelity Matters: A frank discussion about ligase fidelity
TypeBlog PostPublishedMarch 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... -
Important Considerations When Using AAVs
TypeBlog PostPublishedJune 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... -
Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering
TypeBlog PostPublishedDec. 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... -
Antibodies 101: Designing Your First Flow Panel
TypeBlog PostPublishedMay 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 ... -
Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein
TypeBlog PostPublishedJune 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... -
Addgene’s First AAV Data Hub Challenge
TypeBlog PostPublishedSept. 20, 2022, 1:15 p.m....Challenge, now taking place Dec. 6, 2022! Share your AAV data, connect with other researchers working with AAV... -
Suggest A Plasmid You'd Like to Find at Addgene
TypeBlog PostPublishedMarch 16, 2016, 5:30 p.m....community, we’re asking you for plasmid suggestions. Can’t find a plasmid you’re looking for in the repository... -
Participate in the Pets of Addgene Scavenger Hunt
TypeBlog PostPublishedApril 1, 2021, 1:15 p.m....’s Day on the Addgene blog. We’ve been mostly working remotely for the last year to give our lab and ...in the raffle, you will receive an email from us asking for your mailing address and T shirt size. You...