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We narrowed to 846 results for: ELL

Showing: 401 - 420 of 846 results

  1. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ... want to make sure your cells have first reached an appropriate density. Cells in the exponential growth...collecting dead cells, and, conversely, you cannot induce too early as there are not enough cells in the culture... purposes, these E. coli strains are not usually well suited for recombinant protein expression. Many ...host protein, using up valuable resources in the cell thus leading to slowed growth. Additionally, some... promoter. Although the DE3/T7 RNAP system works well for most experiments, the lac promoter can “leak...such as TB or 2XYT is optimal because of the high cell-density they support; however, minimal media supplemented...

  2. Fluorescent Proteins 101: History of Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 7, 2017, 1:58 p.m.
    ...crystal jellyfish Aequorea victoria (Figure 1) in molecular detail. On the underside of the “umbrella” of ... chemiluminescent proteins, like luciferases, as well as fluorescent ones, like Green Fluorescent Protein...green glimmer, Shimomura collected many, many jellyfish specimens from Puget Sound in the Pacific Ocean...he was able to isolate two proteins from the jellyfish’s photoorgans; the first, which he called aequorin... could be used to measure gene transcription, as well as to track protein localization. And as it turns..., another part of the fluorescent protein puzzle fell into place, as they, taken alongside the insights...

  3. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    Published
    June 17, 2025, 1:15 p.m.
    ...transfection or viral delivery of a plasmid. RNAi is an umbrella term for a number of different applications that...miRNA and piRNA can be found endogenously in many cell types, while shRNA and siRNA are synthetic constructs...methods. shRNA processing Following delivery into cells using viral vectors or plasmids, shRNAs hijack the...shRNAs is to generate stable knockout or knockdown cell lines. In fact, it was the go-to choice for many...shRNAs are a quick way to create stable knockdown cells lines, and you can usually find validated shRNA ...Resources References Bartel, D. P. (2004). MicroRNAs. Cell, 116(2), 281–297. https://doi.org/10.1016/s0092-...Montgomery, M. K., Kostas, S. A., Driver, S. E., & Mello, C. C. (1998). Potent and specific genetic interference...

  4. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    Published
    May 4, 2023, 1 p.m.
    ...knock out, etc. – CRISPR can be used to generate cell line or animal models which have the precise disease...Since CRISPR has been adapted for use in primary cells, many tissue types, and across species, it enables... phenotype. CRISPR can also be used to generate cell line knock-ins of patient mutations which are variants...CRISPR has revolutionized the generation of knockout cell lines which has made this control step extremely...in a cancerous background and treat the parental cell line and the X knockout with your drug(s). If a ...research field! References and Resources References Fellmann, C., et al., Cornerstones of CRISPR-Cas in drug...the Addgene blog CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit Disease Out of the...

  5. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

    Type
    Blog Post
    Published
    Jan. 21, 2020, 2:30 p.m.
    ... disease, cell type or drug treatment can browse through screens performed in relevant cell lines or conditions... currently houses over 890 CRISPR screens in 629 cell lines, each manually curated and annotated from ...feasibility and power of CRISPR-based library screens in cell lines from humans and other species. By accurately... screens performed in 26 different breast cancer cell lines. Screenshot updated 2024. Each screen ...screen is captured by BioGRID curators, including the cell line/tissue type, enzyme system, sgRNA library and...example phenotype screens with read-outs other than cell viability. The following video provides an in-depth...

  6. Addgene at the ASM Microbe 2016 Conference

    Type
    Blog Post
    Published
    July 7, 2016, 2:30 p.m.
    ...plasmid sharing services, as well as #happydepositors who stopped by to say hello. Needless to say we all ...booth. As well as being the Director of the Great Lakes Bioenergy Research Center and a fellow Rhodobacter... plenty of geeky fun to be had at ASM Microbe as well as the serious business of science. Two symposia... work this Broad Institute lab is doing creating well-optimized CRISPR screening technology. Addgene got...making popular screening libraries like Brie and Brunello available to the scientific community. However...

  7. Antibodies 101: Epitope Availability

    Type
    Blog Post
    Published
    Jan. 11, 2022, 2 p.m.
    ... have been extracted from a cell, instead of when it is attached to a cell membrane.  Picking the Right...even in that mythical place where antibodies are well-organized, documented, and easy to find (though ...partners, the epitopes available for binding change as well. If this change hides the epitope that the antibodies...with native proteins instead of denatured ones - well, don’t relax just yet. Proteins often change shapes...epitopes that are available on the outside of the cell. The same protein may have different epitopes available...need to validate on denatured proteins or on live cells. But for others, you may need to validate that it...

  8. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    Published
    April 25, 2019, 3:01 p.m.
    ... a look! mNeonGreen: a yellow-green fluorescent protein mNeonGreen is a yellow-green fluorescent protein...protein that was derived from the tetrameric yellow fluorescent protein (LanYFP) from the marine invertebrate...was reported as the brightest monomeric green or yellow fluorescent protein at the time. It is 1.5 to 3...mNeonGreen wavelengths lie between the green and yellow fluorescent protein wavelengths, it can be imaged...with standard green fluorescent protein filters or yellow fluorescent protein filters, with no or minimal...a FRET donor to mCitrine and mOrange or for live cell imaging of fusions and FRET reporters. mTFP1 is ...

  9. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    Published
    Feb. 8, 2024, 2:15 p.m.
    ...millions of cells. For every individual sgRNA, you recover a minimum of 1,000 targeted cells to sequence...trying to generate a knockout cell line, with the goal of isolating single cell clones to use for multiple...generate a ssDNA break which can easily be repaired by cells with a near zero likelihood of mutation arising.... a level of, say 5%, then the majority of single cell editing events will be on target. Thus, if you isolate...example, if your goal is to generate a knock-out cell line to perform downstream characterization of, ...experiments on is a good way to control for off targets as well as clonal differences. If you are performing CRISPR...

  10. A Control for All Seasons

    Type
    Blog Post
    Published
    Oct. 5, 2023, 1:15 p.m.
    ...to detect dead or dying cells so that you can base your analysis on living cells only (for example, propidium...Fluorescence Microscopy.” Journal of Cell Biology, vol. 185, no. 7, Rockefeller University Press, June 2009, ... positive control group, with an alternative but well-studied kinase known to prevent tau hyperphosphorylation...interactions will be observable in preimmune serum ‌as well as with an isotype-matched control.      ...This tends to be most problematic when staining cells or tissues that are the same species as your primary...propidium iodide only stains dead cells). Compensation controls address the spectral overlap that occurs...

  11. Scientists Map the SARS-CoV-2-Human Interaction Network

    Type
    Blog Post
    Published
    April 9, 2020, 1:15 p.m.
    ... used HEK293T cells in our first study, but we're planning to soon expand to other cell types, including...26 of the 29 SARS-CoV-2 viral proteins in human cells and used these viral “bait” proteins to fish for...human proteins that the virus interacts with in cells. Because viruses rely on the help of human proteins...monkeywrench into the virus’ hijacking of human cells. Even better, more than half the drugs hitting these...they have an IRES-Puro marker to facilitate stable cell clone selection. Expression is driven by a fairly... as the EU, Russia, India and Saudi Arabia -- as well as closer to home in the US and Canada. In fact,...

  12. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    Published
    July 17, 2018, 12:23 p.m.
    ... of human stem cells when students are not even allowed to manipulate prokaryotic cells? Especially, if...should be “brains-on”, but as much as I try being compelling, interactive, and application-driven to boost...where you need to apply basic molecular biology, cell biology, microbiology, genetic engineering and strategic...could be used to genetically manipulate human stem cells. Then they had 10 minutes to expose their strategy...construct and explain why they selected a certain target cell population. Who is the winner? Ultimately, society...gene therapy in the following lessons. And let me tell you more, they had brilliant ideas on how to solve...

  13. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    Published
    April 7, 2022, 1:15 p.m.
    ... glutamate release from neurons and other brain cells in vivo. Glutamate plays a variety of roles in synaptic...(SF-Azurite-iGluSnFR), green (SF-iGluSnFR), and yellow (SF-Venus-iGluSnFR). iGluSnFR3: Improved Glutamate...photostability, quantum yield, and expression. Create yellow (v82) and lime green (v857) variants differing ... versatility through two different variants The yellow (v82) and lime green (v857) versions can be summarized...sensor to the membrane and displays it on the extracellular side. In addition to the original PDGFR TMD,...promoter, FLEXed for expression in Cre-expressing cells hSynap: Human Synapsin-1 promoter. Good for neuronal...promoter, FLEXed for expression in Cre-expressing cells. hSynap-FLP: Synapsin-1 promoter, FLP-dependent ...

  14. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    Published
    July 27, 2017, 1:11 p.m.
    ...Awardees: Xin Zhou and Fatima Enam. There were many excellent applicants but Xin and Fatima stood out for their... is currently a graduate student in Thomas J. Mansell’s lab at Iowa State University where she is developing...reliably detect HMOs. Fatima’s first design is a whole cell biosensor with two components - an enzyme that breaks...proposes using this sensor to detect HMOs found on cellular surfaces. After incubating the sensor with bacteria...binding results in an increase in fluorescence of cells that actively produce the HMOs. Fatima has demonstrated...work at her home town in Bangladesh where it was well received. In addition to her research endeavors,...out Fatima's recent publication on her HMO work in Cell Chemical Biology! Additional Resources on the Addgene...

  15. 5 Great Apps for Lab Life

    Type
    Blog Post
    Published
    Sept. 22, 2016, 2:30 p.m.
    ...’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn a new...spectrometry app. I digress. When I started learning proper cell culture technique, I tried to find an app that could...calculations. This is how I came across the Gibco Cell Culture app. Gibco organizes its app into four categories...tools are a lifesaver when your research requires cell culture and include a counter, lab timer, dilution...educational section. This section holds the Gibco Cell culture manual and basic instructional videos. The... Overall, it’s a great tool for someone starting cell culture work. 3. NEB Tools by New England BioLabs...alternative, similar apps: Findings Lab Notebook Cell Culture Split Calculator by DRDP Restriction Enzyme...

  16. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    Published
    Sept. 8, 2015, 2:30 p.m.
    ...Marcotte EM, Voigt CA, Ellington AD. Cell. 2009 Jun 26;137(7):1272-81. doi: 10.1016/j.cell.2009.04.048. PubMed...with sophisticated gene expression controls. Most cell types do not respond to low-intensity light, so ...bacterial camera that prints a chemical image, as well as a genetic method for the computational problem... are compatible with many strains of E. coli, as well as with other plasmid-based systems, which will ...ZB, Lavery LA, Levy M, Davidson EA, Scouras A, Ellington AD, Marcotte EM, Voigt CA. Nature. 2005 Nov 24...

  17. Advice for Scientists Starting a Lab

    Type
    Blog Post
    Published
    April 29, 2014, 1:43 p.m.
    ...carefully select first hires who are well-suited, productive, and fit well with your goals. These first hires...and foster an effective working environment? Tom Ellis was offered a start-up package that included a postdoc... most surprising thing about becoming a PI? Tom Ellis has a one word answer: “Email. Nothing prepared ...differently or the same as your adviser did? Tom Ellis merges the influence from his two PhD advisers who...our latest results as soon as the gels ran.” Dr. Ellis tries to lead his team as a scientists and a friend... lab!”   Acknowledgments Thank you to Dr. Tom Ellis (Imperial College London), Dr. Connie Cepko (Harvard...

  18. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...bacteria, so it can be transformed as-is into cells. The cells then get to work fixing up the “damage” and...nicked plasmid together long enough to get it into cells and let them do the repair work. The key is T4 DNA... update, courtesy of Addgene depositor Stephen Elledge. His new method, named sequence- and ligation-independent...assembled in a unique order with no cloning scars. The Elledge Lab showed that a five-piece assembly reaction ...vitro to generate recombinant DNA via SLIC. Li MZ, Elledge SJ. Nat Methods. 2007 Mar;4(3):251-6. Epub 2007...sequence- and ligation-independent cloning. Li MZ, Elledge SJ. Methods Mol Biol. 2012;852:51-9. doi: 10.1007...

  19. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    Published
    May 3, 2018, 1:48 p.m.
    ...editor in mammalian cells Not all Cas13d proteins are functional in mammalian cells, but Konermann et al... Ruminococcus flavefaciens (nicknamed CasRx), as well as EsCas13d. Find the Yan et al. plasmids here ...make it possible to target essentially any RNA in a cell. Since catalytically dead dCasRx maintains its RNA-binding...natural diversity of CRISPR systems, only time will tell how large the genome and transcriptome engineering... with RNA-Targeting Type VI-D CRISPR Effectors.” Cell (2018) pii: S0092-8674(18)30207-1. PubMed PMID: ... a WYL-Domain-Containing Accessory Protein.” Mol Cell. (2018) pii: S1097-2765(18)30173-4. PubMed PMID:...

  20. A Deep Dive into BioBuilder

    Type
    Blog Post
    Published
    Oct. 11, 2017, 1:44 p.m.
    ...want to learn more about Synthetic Biology. Can you tell me where to start?” is a question more and more ...2011, BioBuilder is the brainchild of Dr. Natalie Kuldell from MIT’s Department of Biological Engineering...from her 20 years of college teaching experience, Kuldell sought to create a program that would capitalize...contribute as part of a team.” As a seasoned educator, Kuldell also knew that the best days in the classroom are...plan or outstanding test scores.  As a result, Kuldell launched the BioBuilder Educational Foundation ...of Professor Richard Murray. Murray’s lab uses cellular extracts to perform transcription and translation...develop and test hypotheses. Students then use cellular extracts as a means to rapidly prototype DNA circuits...
Showing: 401 - 420 of 846 results