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We narrowed to 1,010 results for: PLE

Showing: 461 - 480 of 1010 results

  1. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    ...This second plasmid saves you a couple of weeks. CRISPR saves you a couple of months. It’s higher throughput...system work? Mendenhall: The system is really quite simple but it offers improvements in efficiency and throughput...right thing to do to put these out there. It’s a simple solution. It would be silly not to share it as ...Since the Addgene page went up last month, over 340 people have signed up to receive alerts when plasmids ...additional tips, tricks, or words of advice for people who may want to apply this tagging system to their...

  2. From Lab Bench to Leadership: Transitioning into Management as a PhD

    Type
    Blog Post
    ...motivations, strengths, and learning styles. For example, during my time as a lab scientist, I worked with... management. Invest time in understanding key principles through books, workshops, and online courses ...foster a cohesive, productive environment. For example, in my current role managing a cross-functional...facilitated a team meeting to highlight their complementary strengths. By encouraging alignment on goals... while changing industries and positions. Some people discover they truly enjoy management and choose...successfully transitioned to industry after his PhD and a couple of postdocs focusing on molecular biology and metabolism...

  3. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ... cell research has prompted the development of multiple virus packaging methods. Across these methods,...transfection step of the viral packaging cell line. For example, Phoenix, a second generation γ-retrovirus packaging... to enter the nucleus through the nuclear pore complex. This feature may be useful for the transduction... the components found within the backbone. For example: pLXSN has the wild type LTR, an MCS for cloning...sequence. This post was written and updated by multiple Addgenies. It was originally published in 2018...  Multiple Authors...

  4. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

    Type
    Blog Post
    ...ever like to write about a recent plasmid deposit please sign up here.  Listen to the hot plasmids episode...to physically set up in the lab, RNAi is often completely ineffective or results in low penetrant phenotypes...low-cost, cell-free detection assay that can be completed without special equipment. What’s more, detection...encodes the LacZ𝛼 protein. Once expressed, LacZ𝛼 complexes with LacZ𝜔 and forms an active LacZ tetramer....red-b-D-galactopyranoside on the assay paper turning it purple, allowing for easy visual detection of the presence...modifying the sequence of the toehold switch. For example, Green has also developed toehold switches to detect...

  5. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    ...working to visualize multiprotein complexes.  Fig. 2. Multiplex immunolabeling with subclass-switched...easily be engineered. Like humans, mice express multiple subclasses of IgG antibody. Broad range secondary...subclass specific secondary antibodies to detect multiple targets with different mAbs in the same experiment...secondary antibodies and subclassed switched R-mAbs, multiple targets could now be visualized in a single experiment...recombinant monoclonal antibodies for enhanced multiplex immunolabeling of brain. Elife. 2019 Jan 22;8:...

  6. Sharing is Caring: But How to Distribute Open Hardware?

    Type
    Blog Post
    ...achievement, the complexity of the device increases, as does the price. With this increased complexity comes the...very difficult to adapt the device to one's own samples and limits the use of microscopy in open science...teleporter," since a part developed in Germany, for example, can also be printed quickly in Madagascar. Platforms...a move towards more open research. The EU, for example, is calling for the opening of sources within the...hardware.    Conclusion Addgene is a great example of how sharing of data and molecular tools can ...

  7. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    ... range from a defined pathway or reaction to a complex event such as cancer or even aging. For both a ...differences in these approaches can be found. As an example, say you are considering either a cellular screen...libraries targeting all known genes. This is a perfect example of an unbiased, reverse screen – all known genes...possessing a specific mutation or phenotype. For example, a drug screen may be performed on a cell line ...patients may respond to drug treatment. Another example is using a mouse model of a particular disease ...that do not affect the function of interest. An example where a selection would work is if you are trying...

  8. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ...fluorescent proteins can be directly compared. An example of a photostability measurement of various cyan...results in a loss of fluorescence, which, in its simplest form, can be described by a mono-exponential decay...intensity decay of a fluorophore does not follow a simple mono-exponential decay (Shaner et al, 2008; Bindels...applications in which far less power is used. For example you might find that you can get useful data from... diffusion, the proteins are (i) trapped in microdroplets in a water/oil emulsion, (ii) embedded in a ...fluorescent protein. In figure 1 we provide an example of such a comparison between cyan fluorescent proteins...

  9. Antibodies 101: Conjugation

    Type
    Blog Post
    ...antibodies present throughout our bodies carry out plenty of roles just the way they are, the research antibodies... the visible spectrum, so they are great for multiplexing and can be used in a variety of assay types....detection of low abundance targets with significant multiplexing potential. You will find antibody-oligonucleotide...Fc-binding domain (Figure 1).  Figure 1: Examples of sites on antibodies used for conjugation. 1...from antibody to antibody or batch to batch. For example, take lysine conjugation, which is commonly used...reaction or the functionality of the conjugate. For example, Addgene’s antibodies are provided in a buffer ...

  10. To Each HIS Own

    Type
    Blog Post
    ...   Fig. 1: A sample containing a polyhistidine tagged protein of interest (purple) and several contaminating...strategy a single protein can be isolated from a complex mix using affinity chromatography against an amino...metal ions are immobilized on a bead or resin and a sample containing the protein of interest flowed through...Ni2+-containing resin. The protein of interest (purple) binds to the Ni2+ and is immobilized along with...tags are also recommended if you need a very pure sample. Because binding strength is proportional to the...

  11. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    ...listen quietly as people discuss their experiences. If you’re hearing multiple people with similar misconceptions... that a lot of science communication is helping people understand new concepts. You hopefully are aware...necessary” detail after “necessary” detail and end up completely overwhelming your audience - like the professor...group was just really tired and needed a slower, simpler presentation than I had planned; other times students...

  12. Science communication: 9 Strategies to Get Your Foot in the Door

    Type
    Blog Post
    ...public, for example at an anniversary or for Girls’ Day, offer guided tours to show people where the lab...into science education Do you aim to reach young people? Try networking with your local education community...Trust blog or the John Innes Student Voice are two examples. Many blogs like Plant Science today, Soapbox ...meetings. 8) Training – Get a formal education Some people get into science communication by taking a media...them to be the “be-all-and-end-all”. You can, for example, get an MSc Science Communication or MSc Science...

  13. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ... features of the gene intact. Here, we show an example gene targeted for tagging with GFP at the C terminus...interest. For a C-terminal tag, as shown in the example above, we inserted a linker and GFP tag at the ...the GFP tag precisely where we need it, in this example, at the C-terminus of the gene of interest. To ...CRISPR/Cas9 (Cas9 protein and the crRNA-tracrRNA complex) and the donor plasmid into the cells we use electroporation... Institute for Cell Science Plasmid Collection, please visit the Cell Catalog on the Allen Cell Explorer...hiPS cell lines generated using these plasmids, please visit the Cell Catalog on the Allen Cell Explorer...

  14. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...Having never been to China myself, I was pleased to be able to participate when the organizers of the...Lowe lab have allowed them to begin understanding complex regulation of and by tRNAs and tRNA-derived RNAs...long hypothesized that life started with RNA and a simple cell membrane. His recent work shows that they ...replication with no proteins). The chemistry is complex and this work requires an interdisciplinary team... they do it. Of course, No conference would be complete without CRISPR Michael Terns from of the University...structural and functional component of the ribosome complex and Dr. Steitz quipped that RNA in the ribosome...

  15. Tips for Technical Support Calls

    Type
    Blog Post
    ...buried beneath someone's lunch. Many times, I’m pleasantly surprised to discover that the most crucial information...from you and it’s wrong. Can you help us fix it? Please address as soon as possible!” and I always begin...your concern is based on data you have produced, please share the data. Maybe not all, but many bio companies...helping us understand the nature of your concern. Please be courteous. We understand how exasperating it...time and money on a potentially faulty product. Please remember that the person at the other end of that..., let your customer service agent know (nicely, please!). We can’t improve without feedback. And if you...

  16. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    ...toxins are the clearest example of a plasmid benefiting its host. For example, cells with a penicillin...Shareck et al., 2004). Plasmid categorization is complex Plasmid classification has shifted over the years...Plasmids may also have more than one function: for example, Gateway cloning relies on genes found on an E....plasmids without clear analogues in nature. For example, helper plasmids were developed to replace helper... Multiple Authors...

  17. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    ... parents. Here’s how to cross mice, using a simple example with the goal is to generate mice that have...genome, they are not commonly used to introduce multiple mutations. This is because as more mutations are...challenge that may arise when crossing mice with multiple mutations is if the mutations are on the same ...expressing strains Cre-lox strains are a common example of mice that are derived from crosses. In Part ...With this two-part guide to mouse modeling now complete, you should be prepared to plan an experimental...

  18. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ...getting-started guide for BLAST, which includes a simple explanation of the different BLAST programs, databases...consistently run our blastx searches with the “Low complexity regions” filter unchecked so that these regions...benefit is that blastp searches do not filter low complexity regions by default, meaning that you do not have...Clustal is our frequent choice for pairwise or multiple sequence alignments of nucleotide or protein sequences... sequences. We also use COBALT for aligning multiple protein sequences, particularly for comparing different...//www.ebi.ac.uk/Tools/webservices/#multiple_sequence_alignment_msa http://www.ebi.ac.uk/Tools/webservices...

  19. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...allowing very low background level of fluorescence. Examples for such photoactivatable or photoconvertible ... and their emission is green or red light. For example, T-Sapphire, LSSmOrange, and LSSmKate. Fluorescent...sensor), and iGluSnFr (glutamate sensor). More examples of these biosensors can be found at Addgene. ...also is also called Bimolecular Fluorescence Complementation (BiFC). 8 points to keep in mind when choosing...system can excite, and detect the emission. For example, if your microscope has only two lasers, at 488nm...minutes after it is translated to a few hours. For example, superfolder GFP (sfGFP) and mNeonGFP can fold ...

  20. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...strengthen your results. Enrichment or depletion of multiple gRNAs targeting the same gene can be strong evidence...jar of jellybeans, a pooled CRISPR library is a complex mixture. Pooled libraries are composed of many ...sorting and screened for a particular phenotype. For example, mutant cells can be used in drug screens to identify.... Loss of representation due to enrichment or depletion of specific gRNAs can lead to skewed results. ...any experiment, the use of appropriate controls, multiple replicates and several cell types can strengthen...
Showing: 461 - 480 of 1010 results