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We narrowed to 672 results for: abo.1

Showing: 41 - 60 of 672 results

  1. Tetracycline Inducible Expression

    Type
    Collection
    ...Element On or off PI 26429 pLenti CMV rtTA3 Blast (w756-1) Lentiviral reverse tetracycline-controlled transactivator... or shRNA; see article for detailed information about cloning and additional plasmids tetR-KRAB On Aebischer...

  2. CRISPR References and Information

    Type
    Collection
    ..., 2.4 MB Zhang GeCKO pooled library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9...screening. CRISPR eBook Addgene's downloadable eBook about all things CRISPR. Addgene CRISPR blogs CRISPR tips...Developed by the Fredrik Wermeling Lab . Read more about it on the Addgene blog . CRISPOR (Link opens in ... Lab and maintained by the Frederick National Laboratory at the National Cancer Institute. Cas-OFFinder...maintained by the Jennifer Doudna Lab . Read more about it on the Addgene blog . CRISPRflydesign (Link opens...

  3. Antibody Guide

    Type
    Collection
    ...and placed in a buffer. Antibody Structure Figure 1: Structure of an antibody A standard antibody is made... together to form a “Y” shape, as shown in Figure 1. The two arms of the Y structure are responsible for...are often used in clinical applications. Diabodies - Diabodies contain two Fab fragments, each recognizing...cells isolated from animals immunized as described above are used to create hybridomas, which produce large...different epitope, bound by short peptide linkers. Diabodies can be used for assembling protein nanostructures... Includes: Flow cytometry Read on to learn more about the applications common to each category, including...controls, and any special considerations to think about. For step-by-step instructions of many of these ...

  4. COVID-19 Resources

    Type
    Collection
    ...mediating the entry of SARS-CoV and SARS-CoV-2 viruses. ( 1 ) TMPRSS2 - a serine protease that primes the SARS-CoV...lightning speed to elucidate as much as they can about this novel virus. It is now known that SARS-CoV-...and open-access SARS-CoV-2 detection assay for laboratory and home testing. Kellner, et al. bioRxiv 2020.06.23.166397...

  5. Who Gives a Tweet? 9 Facts About Scientists on Twitter

    Type
    Blog Post
    ...2017. 1. Among scientists, historians have the most Twitter users. Biologists rank #8. Figure 1: Highly...that these results could be biased for two reasons: 1) they are based on the US Occupational Employment ...female to male = 0.629 (the closer this ratio is to 1, the closer males and females are to parity - the ... authorship in the United States (0.428). 4. The #1 (re)tweeted domain by scientists was Instagram Social...links to research that isn’t behind paywalls. 5. The #1 (re)tweeted science domain by scientists was Nature...use Twitter professionally to network, learn more about alternative careers for scientists, and share cool....” This paper addresses the following questions about scientists on Twitter: who are they? What do they...

  6. Retrograde AAV viral preps

    Type
    Collection
    ...NLS-mRuby2 Control Gradinaru 83900 pAAV-mDlx-GFP-Fishell-1 Dlx GFP Control Fishell 83895 pAAV-hDlx-Flex-GFP-Fishell...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...

  7. Bacterial Expression Systems

    Type
    Collection
    ...by red light. pCph8 50552 pLtetO-1 Anhydrotetracycline Jeffrey Tabor Anhydrotetracycline inducible expression...in vitro ? These, in addition to many questions about bacteria themselves, can be answered using model...overexpressing or knocking down a gene can tell you much about its function and/or alter cellular activity in a...basic biology of a bacterial gene or redirecting metabolic pathways to produce a therapeutic compound, there...once you get it right, you can learn many things about your protein’s function, structure, and interactions...molecules, light, temperature, or even intracellular metabolite levels. Different plasmids and, often times, ...

  8. Luciferase Plasmid Collection

    Type
    Collection
    ... Eric Campeau 21474 pLenti CMV V5-LUC Blast (w567-1) Firefly CMV Lentiviral expression of firefly luciferase...Plasmids are Gateway cloning compatible. Learn more about DULIP in our 27 Hot Plasmids from 2016 blog post...circularly permuted Venus fluorophore. Learn more about pHlash sensor in our 28 Hot Plasmid Technologies... with the original STARR-seq method. Learn more about STARR-seq in our 27 Hot Plasmids from 2016 blog ...

  9. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...plasma HIV-1, improving life expectancy and quality for HIV-1 patients. However, ART is not an HIV-1 cure. ...activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that can produce HIV-1 toxic...with HIV-1. For two HIV-1 strains, Cas9/gRNA lentiviral expression significantly reduced the HIV-1 copy number...currently infected with the lentivirus HIV-1. Today, HIV-1 can be controlled with antiviral therapies ...HIV-1 transcription and destroy infected cells, the other using wild-type Cas9 to remove the HIV-1 genome...take on HIV-1 in vitro, and what obstacles must be overcome for clinical success. ART and HIV-1 reservoirs...Cutting HIV-1 out of the genome with CRISPR/Cas9 Before the introduction of CRISPR, HIV-1 genome editing...

  10. Revamp Your Lab Meetings With Creative Virtual Collaboration

    Type
    Blog Post
    ...  Figure 1: Virtual lab meeting? No problem! You can still creatively collaborate and brainstorm... on it – so it is important to take breaks. After 1 hour (or another designated time) of meeting, someone...else to focus on. Use collaborative cloud apps It is a good idea to use collaborative cloud tools such as...creative collaboration in the workplace. There are many other ways to encourage collaborations remotely...serve as a springboard for future discussion. Collaboration and creativity are pivotal in writing publications...Vicens et al., 2007) geared towards enhancing collaborative writing and this recent article published in...approaches/tools we have to foster creative collaboration remotely in science are endless. Let’s adapt...

  11. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...are based on the Epstein-Barr Nuclear Antigen-1 (oriP-EBNA-1) system where each vector contains a viral ...replication, OriP and the EBNA-1 sequence that encodes for the DNA binding protein EBNA-1 [7]. These two sequences...extra plasmid, pCXWB-EBNA-1 is used in the reprogramming cocktail. pCXWB-EBNA-1 lacks oriP and is unable...expressing additional EBNA-1 transiently. The transiently expressed EBNA-1 increases protein expression...pCXLE vectors listed below in Table 1 are available at Addgene. Table 1: List of pCXLE episomal plasmids,...obtained and the subsequent reprogramming success [1]. Fibroblasts need to be cultured and expanded in ...reprogramming factor cocktail, and viral protein EBNA-1. Methods for reprogramming hiPSCs from PBMCs While...

  12. Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond

    Type
    Blog Post
    ... Figure 1: Crowdfight is a community of scientists who believe in the power of collaboration and join...When a researcher needs a collaborator for a specific task, Crowdfight will: 1) Find an expert with the...our blog post about remote collaborations Read our Transferable Skills Guide article about external collaborations...needed. In the example above, for example, we fostered a formal scientific collaboration between two research...project. We think that these collaborations add enormous value. A discussion about a protocol can save weeks..., the researchers did not have access to a P3 laboratory (BSL-3), a high-security facility needed to perform...approach was useful in humans. Then, one of them heard about our online platform, called Crowdfight, and filed...

  13. DIY DNA Ladders from Penn State University

    Type
    Blog Post
    ...Weight Markers. Scientific Reports,7(1). doi:10.1038/s41598-017-02693-1. PubMed PMID: 28550309. PubMed Central...plasmids that can be used to make inexpensive 100 bp or 1 kb DNA molecular weight ladders were recently deposited... PstI or EcoRV, these plasmids generate 100 bp or 1 kb DNA ladders, respectively. Unlike many commercially... gels. Get Tips on Verifying Your Plasmid Table 1: Comparing Typical Comericial Ladders with the PSU...$10 to make 1000 lanes worth of the 100 bp or the 1 kb ladder. Using a commercial plasmid prep kit increases...Learn How to Analyze a Restriction Digest References 1. Henrici, R. C., Pecen, T. J., Johnston, J. L., & ... Box Find 5 Smart Phone Apps for Lab Life Learn About the Evolution of Lab Techniques Resources on Addgene.org...

  14. Adenovirus Guide

    Type
    Guide
    ...to an antigen. AdEasier-1 cells BJ5183 E. coli cells containing the pAdEasy-1 packaging plasmid. Adenovirus...icosahedral capsids and build new virions. Figure 1: Wild-type adenovirus and its genome. Genes over or... This construct is then transformed into AdEasier-1 cells (Addgene #16399) , which are BJ5183 E. coli ... viruses. The adenoviral backbone plasmid pAdEasy-1 (Addgene #16400) is suitable for most purposes. For...production system. Figure is based on the use of pAdEasy-1. Created with BioRender.com. AdMax™ AdMax™ is another...landscape . Signal Transduction and Targeted Therapy, 6 (1), 53. https://doi.org/10.1038/s41392-021-00487-6 (...as a Zika virus vaccine . Scientific Reports, 10 (1), 3613. https://doi.org/10.1038/s41598-020-60238-5...

  15. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...Table 1 below outlines a few of the more common genetic changes found in E. coli strains. Table 1: Common...by tyrosine insertion λ-thi-1 or thi1 Mutation in thiamine metabolism Requires exogenous thiamine for... all based on E. coli K-12 and are considered BSL-1. Table 2: Lab strains of E. coli Strain Natural...plasmids, blue/white screening. F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF...) recA13 leuB6 ara-14 proA2 lacY1 galK2 xyl-5 mtl-1 rpsL20(SmR) glnV44 λ- JM109   General cloning and...strain for cloning repetitive DNA. endA1 glnV44 thi-1 relA1 gyrA96 recA1 mcrB+ Δ(lac-proAB) e14- [F' traD36...of an E. coli K-12 strain. F- λ- ilvG- rfb-50 rph-1 NEB Stable   For cloning into and storage of lentiviral...

  16. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    .... The entire process from start to finish took about 1 month. Cas9 delivery Overview: Before any editing...blotting is a loading control. Pro-Tips: 1. It’s all about the MOI. In retrospect I used a way too high...to knockout BRAF in a human cancer cell line in about 1 month, which I’d say is pretty amazing. Despite...lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells were then used to generate a monoclonal...rest of the cells were frozen down. Figure 1: Cas9 expression in A549 cells transduced with lentiCas9... pool had wildly variable Cas9 expression (Figure 1). This is a problem because Cas9 expression affects...cells were transduced (MOI = 37) and selected with 1 µg/mL blasticidin for 9 days. Single cells from a ...

  17. Great Results Start with Great Standard Curves

    Type
    Blog Post
    ...    Figure 1: A 1 mg/mL control is diluted in a series ranging from 1:2 to 1:32. Each individual...example, Figure 1 depicts a 2-fold dilution series. Each step of the series dilutes by 1:2 for a series...series that ranges from 1:2–1:32. The specific dilution series used will depend on the expected concentration... points fit the trendline. R2 ranges from 0 to 1 with 1 being a perfect fit and 0 indicating that there...individual step of the series is consistent at 1:2. When preparing a dilution series, use a new pipette tip for...by replicate values provides useful information about the accuracy of the curve, which we will discuss...many scientific applications users aim for an R2 above 0.95.    Additional considerations When running ...

  18. Lentiviral Vector Guide

    Type
    Guide
    ...production in the laboratory, and many have been removed or mutated for increased safety. Figure 1: Wild-type...typically ranges from 8–10 kb, encoded on RNA (Figure 1). This RNA is reverse transcribed into the provirus...genes (for example, vif , vpr , vpu , and nef for HIV-1). The LTRs flank all of these genes, and anything ...transgene and LTRs (chimeric 5’ LTR) Packaging plasmid 1 — contains gag and pol Packaging plasmid 2 — contains...and third-generation systems, please refer to Table 1. Figure 4: Third-generation lentiviral plasmids. Created...heterologous enhancer/promoter such as CMV or RSV Table 1. Summary of key differences between second- and third-generation...More Lentiviral Vector topics Addgene Protocols pLKO.1 - TRC Cloning Vector Protocols Lentiviral Vector Production...

  19. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ... chromosome, start, and stop. For instance (chr3, 1, 10) indicates a stretch of DNA at the very beginning...genetic elements like protein, RNA, and DNA (Table 1). As with GPS in the real world, coordinates alone...genomic coordinates into meaningful experiments. Table 1: An emerging family of "GPS for the genome" techniques...enhancers with the help of genome math. Figure 1: Genome math illustrated. Intersect (left) and set...histones with “GPS for proteins,” ChIP-seq from Table 1. Because of DNA’s 3D geometry and the chemical properties...experiment to identify the critical NANOG binding sites: 1. Find Enhancers that have NANOG binding sites 2. Design... In order to denote the NANOG enhancers from step 1 with genome math, we’ll need a bit of shorthand from...

  20. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    ... assembly reaction** 1 5 min., 37 ºC (2.5 μl) 687 100% 274,200 2,742,000 1 60 min., 37 ºC (2.5 μl)...expected complete sequence for the lacI/lacZ genes (1), while sequencing of white colonies showed a mixture...representative transformation plates obtained from 1-, 12- and 24-fragment assemblies of the lacI/lacZ ...lacZ cassette, and illustrates how the volume of the 1 ml outgrowth spread on each transformation plate can...fidelity studies conducted by Potapov et al. at NEB (1, 2). While T4 DNA Ligase, the mainstay of most biotechnological...over 90% for 24-fragment assemblies (Figure 3, Table 1). These improvements (5- to 12-fold increases in the... 16 ºC) x 30 (100 μl) 78 90.7% 783 9,792 Table 1. Yields and fidelities for Golden Gate Assemblies ...
Showing: 41 - 60 of 672 results