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We narrowed to 604 results for: abo.2

Showing: 101 - 120 of 604 results

  1. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...targeting a specific gene within the genome (see Fig. 2). Individual gRNAs are designed in silico using publicly...mouse, human, fly, E. coli, and T. gondii genes.  2) What genetic modification are you trying to make?...CRISPR libraries is a multi-step process (see Fig. 2). In most cases, CRISPR libraries are provided at ... is included on the gRNA-containing plasmid, or a 2-plasmid system in which Cas9 must be delivered separately...uncontrolled cell growth, such as cancer. Figure 2: The CRISPR library must be amplified (A) to be used...preparation from Addgene, you can skip the steps above! Cells are then transduced with lentivirus containing...practices, CRISPR libraries can help you learn a lot about your phenotype of interest. For more detailed information...

  2. Visualizing Your Plasmid Request Data

    Type
    Blog Post
    ...with request data to the dashboard (Fig. 2).     Figure 2: Insert showing the upload function for... the date of downloading the CSV is shown (figure 2) and the dashboard displays the data in a standardized...analyzing and sharing your own depositor metrics!  About the Author Joachim Goedhart is an assistant professor...

  3. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ... of 2 components: 1) an SB transposase, the enzyme required for catalysis of transposition; and 2) a transposon... gene (red) for expressing the transposase enzyme.2.) The transposase enzyme is expressed (red stars) ... 2015 Feb 4. doi: 10.1002/biot.201400821. PubMed. 2. Mates L, et al. Molecular evolution of a novel hyperactive...of DNA that can move around in a genome. In a laboratory setting, transposons can be used to both introduce...

  4. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ...two genes over time, at a range of distances from 2-75 Mbp. Labeling multiple, intrachromosomal loci Building...successfully resolved loci with physical map distances of 2 and 75 Mbp, with the calculated fluorescent distances...established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that they could evaluate the...dCas9 supplied by a separate vector. Figure 2: CRISPRainbow generates 6 unique colors and a white... Find plasmids from this publication at Addgene. 2. Ma, Hanhui, et al. “Multicolor CRISPR labeling of...Additional Resources on the Addgene Blog Read More about Fluorescent Protein-Based Tools Learn Some of the...

  5. Plan Your Experiment

    Type
    Collection
    ...either (1) a lack of gRNA and/or Cas9 expression or (2) a lack of efficient target cleavage in cells expressing...components to make lentiviral particles (for details about lentivirus, see our Lentivirus Guide ) Stable, tunable...when carrying out CRISPR experiments. Read more about how to design your gRNA . Browse Plasmids: gRNAs...to optimize your delivery conditions. Learn more about CRISPR delivery in mammalian systems . Validate ...

  6. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    ... to the lysosome where they are finally degraded (2, 3). Once you've created your fusion protein, it's...examples include the ImageJ plugins JACoP and Coloc 2, BioImageXD, and customized CellProfiler pipelines...colocalization analysis example is given in Figure 2 (adapted from 4) - Dunn et al. 2011 provides an in-depth...Since colocalization relies upon the detection of 2 independent fluorescent signals and their potential...here! References 1. Allen Institute Plasmid Pages 2. Cooper GM. 2000. The Cell: A Molecular Approach. ...the context of the cell. However, much knowledge about functionality can be gained by understanding the...subcellular marker proteins, it’s possible to get an idea about the subcellular structures your protein resides ...

  7. Grad School Advice Part 1: Picking a Lab and a Project

    Type
    Blog Post
    ...Introduction from Tyler 1:13 - 2:02 : Niroshi and Ben introduce themselves 2:03 - 3:22 : How did you end ...excited about coming into lab even when things get tough in the dreaded third year. Find Part 2 Here! ...

  8. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...PubMed Central PMCID: PMC3640491. 2. Grimm, D., et al. "Titration of AAV-2 particles via a novel capsid ELISA...report the physical titer of a viral solution (Figure 2). Because physical titers are used for dosing purposes...mean and how they can be compared.   Figure 2: Label that comes on Addgene’s AAV aliquots. Our titer...of genomes can limit production of recombinant AAV-2." Gene therapy 6.7 (1999). PubMed PMID: 10455443. ...Characterization of a recombinant adeno-associated virus type 2 Reference Standard Material." Human gene therapy 21.10... Addgene.org Find All Available AAV Learn More about Virus Production Find Viral Vector Protocols ...

  9. Common Injection Routes in Mice

    Type
    Blog Post
    ...Location: peritoneal cavity Gauge: 25–27 Volume: < 2–3 ml Skill required: intermediate Intramuscular Intramuscular...Location: back or hind legs Gauge: 25–27 Volume: < 2–3 ml (~1 ml per site) Skill required: beginner Intradermal...Intraperitoneal Scruffing Peritoneal cavity 25–27 < 2–3 mL Intermediate Intramuscular Mechanical restrainer...Back or hind legs; between skin and muscle 25–27 < 2–3 mL (~1 mL per site) Beginner Intradermal Anesthesia...Addgene blog Mouse Modeling Part 1 Mouse Modeling Part 2 Five Popular Model Organisms Additional resources ...Routes of administration. In H. Hedrich (Ed.), The Laboratory Mouse. Academic Press. Turner, P. V., Brabb, ...M. A. (2011). Administration of substances to laboratory animals: routes of administration and factors...

  10. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

    Type
    Blog Post
    .... Diagnostic restriction digests are comprised of 2 separate steps: 1) incubating your DNA with restriction...which cleave the DNA molecules at specific sites and 2) running the reaction on an agarose gel to determine...including a 1.2 kb insert. The plasmid was digested with 2 unique enzymes (HindIII and BamHI) and run on an agarose...to serve as a reference band. The uncut DNA (lane 2) shows 3 possible plasmid conformations, with relaxed...1kb ladder (lane 1) that has bands ranging from about 500bp to 10kb, with the 3.0kb fragment having increased...

  11. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...identification.   Fig. 2: Gel image of Plasmid 61564: pBAMD1-2 (~4.7 kb) digested with SpeI and... at ~3.6 kb and ~1.1 kb (lane 3). Uncut DNA (lane 2) has a band at <3 kb, which corresponds to the supercoiled...formation of plasmid multimers. Cell. 1976 Dec;9(4 PT 2):707-16. doi: 10.1016/0092-8674(76)90134-3. PMID: ...dimers in Escherichia coli K-12. Plasmid. 1989 Mar;21(2):138-41. doi: 10.1016/0147-619x(89)90057-7. PMID: ...Plasmids and Chromosomes. Microbiol Spectr. 2014 Oct;2(5). doi: 10.1128/microbiolspec.PLAS-0025-2014. PMID...Behavior of Plasmid DNA. Microbiol Spectr. 2015 Apr;3(2):10.1128/microbiolspec.PLAS-0036-2014. doi: 10.1128...colonies using one of the detection methods described above can help ensure the isolation of the monomeric version...

  12. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    ...low expression, and everything in between (Figure 2). These smears tend to occur when you are looking ...based on your experiment. You can see from Figure 2 that determining where to place your gate based on...the dot plot would be complicated.  Figure 2: (A) a smeary dot plot showing a range of antibody...plot in Figure 4 compared to the dot plot in Figure 2. Quantification using MFI makes it easier to identify...interest. Created with biorender.com. The plot above is for a smeary expression and is quite different.... Additionally, you can gate off the histograms above, allowing you to visualize your data as a dot plot...

  13. CRISPR Antimicrobials

    Type
    Blog Post
    ...Citorik Plasmids here). In both cases, they used 1-2 CRISPR crRNAs targeting sequences found only in certain...resensitize antibiotic-resistant bacteria (Figure 2). The first phage introduced is lysogenic, integrating...machinery targeting 1) the given resistance gene and 2) a second lytic phage. Bacteria that receive this ...bacteria have previously flourished.   Figure 2: Two-phage strategies to resensitize bacteria to antibiotics...237163. Find plasmids from this paper at Addgene. 2. Bikard, David, et al. “Exploiting CRISPR-Cas nucleases...organizations are working to help educate the public about proper antibiotic usage. Although these steps are...laudable, they likely won’t solve the growing problem. Above all, we need new antimicrobials, ideally ones for...

  14. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    ...scientists are using social media professionally, and 2) I needed to start using Twitter. Sharing Chemistry...but my consistent writing for the Addgene blog (~1-2 blog posts per month) and use of Twitter and LinkedIn...PhD 👩🏾‍🔬💜 (@ThePurplePage) February 13, 2014 2. "The Scientist's Top 10 Guide to Twitter" blog post...you social media can positively impact your career, 2) given you tips for using various social media platforms... advantage. It wasn’t until I wrote a blog post about a research paper that explored how scientists connect...saw a request for an Addgene guest blogger post about colony PCR, I signed up immediately. I had used ...science communication skills, while also learning about new scientific developments and helping scientists...

  15. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...month, from 1/22/20–2/23/20, where there were zero plasmid requests. Afterwards, from 2/24/20–3/15/20 (the...Each paper has between 1–32 plasmids and a total of 2–2,590 requests. We received plasmid requests from ...observed between Jan. 1 and Mar. 1, 2020. Figure 2: A simple approximation of the effect of COVID-19 ...However, it is evident that if we believe that 1/22/20–2/23/20 are the more accurate dates of lockdown in China...in his laboratory. Learn more here. Additional resources on the Addgene blog Learn more about plasmid...

  16. MXS Chaining

    Type
    Blog Post
    ... TagBFP 399nm/ 456nm histone 2B (H2B) Chromatin 2 Cerulean 433nm/ 475nm Lyn-tag (derived from the tyrosine-protein...based (chaining-based) cloning. The modules (Figure 2) are contained in individual plasmids and are each...between various cloning methods are listed in Table 2. For example,  chaining-based cloning methods require...suitable for combining endogenous sequences. Table 2: High Throughput Cloning Strategies Method Technology...PMID: 20205762. PubMed Central PMCID: PMC2822740. 2. Engler C, Kandzia R, Marillonnet S. A one pot, one...Resources on the Addgene Blog Read about Gibson Assembly Learn About Other Plasmid Cloning Techniques Plasmids...

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

    Type
    Blog Post
    ...question: C2c1, C2c2 and C2c3 (short for Class 2, candidate 1, 2, or 3). C2c1 and C2c3 are related to Cpf1,...however cutting efficiency of Cas13a is reduced when 2 mismatches are present. The protospacer flanking sequence...PMC5127784. Find plasmids from this paper at Addgene. 2. East-Seletsky, Alexandra, et al. "Two distinct RNase... and functional characterization of diverse class 2 CRISPR-Cas systems." Molecular cell 60.3 (2015): 385...potential applications of Cas13a given what we know about its structure and function? For starters, Cas13a... Check Out Our CRISPR Featured Topic Page Learn about Anti-CRISPRs Multiplex Genome Editing with Cpf1 ...

  18. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...for controlling neuronal activity, channelrhodopsin-2 (ChR2), a blue light-activated cation channel, from...used in tandem with the opsin.      Figure 2: Diagram of excitatory (ChR2) activation by light ...) domains, phytochrome B (PhyB), and cryptochrome 2 (CRY2) (Zhang & Cui, 2015). When stimulated with light...comparative analysis of microbial opsins. Nature Methods, 9(2), 159–172. https://doi.org/10.1038/nmeth.1808 References...comparative analysis of microbial opsins. Nature Methods, 9(2), 159–172. https://doi.org/10.1038/nmeth.1808 Nagel...Hegemann, P., & Bamberg, E. (2003). Channelrhodopsin-2, a directly light-gated cation-selective membrane ...intracellular signaling pathways. Trends in Biotechnology, 33(2), 92–100. https://doi.org/10.1016/j.tibtech.2014.11.007...

  19. Retrograde AAV viral preps

    Type
    Collection
    ...Calcium sensor Ting 83899 pAAV-mDlx-GCaMP6f-Fishell-2 mDlx GCaMP6f expression under the control of the mDlx...serotype from the Alla Karpova and David Schaffer laboratories. These viral vector preparations were produced...Janelia Research Campus viral project team in collaboration with David Schaffer (University of California...

  20. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...kinetics which are illustrated in figures 2 and 3.  Figure 2: Glutamate titration curves for the different...signals upon sensing glutamate using both 1-photon and 2-photon imaging.   Figure 1: 1P and 2P excitation...adapted from Aggarwal et al…, Supplementary Figure 2.   Improved trafficking and localization through...Additional Resources on the Addgene Blog Learn about other Fluorescent Biosensors A Practical Approach...
Showing: 101 - 120 of 604 results