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We narrowed to 1,009 results for: PLE

Showing: 281 - 300 of 1009 results

  1. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...Plasmid Kit Multiple Lentiviral Expression System Kit (Kit #1000000060) The MuLE (Multiple Lentiviral ...several different viral vectors to choose from, for example gamma-retrovirus, lentivirus, Adenovirus, and Adeno-associated...and lentiviral envelope plasmids are listed here. Please note that both lentiviruses and gamma-retroviruses...combined with high-throughput sequencing to perform multiplexed analysis of cells pre-treated with a compound...Transfer Vectors    Expressing your gene of interest pLenti-puro (Plasmid #39481) You can clone your gene ...We are constantly improving our CRISPR reagents. Please check www.genome-engineering.org for the most up-to-date...system facilitates the simultaneous introduction of multiple genetic alterations into mammalian cells. The ...

  2. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...reducing editing from ~30% in control samples to 0-10% in Acr samples. Acr proteins can also be used to prevent...assembly of the crRNA-Cas complex (Zhu et al., 2019). Inhibiting DNA cleavage Example: AcrIIC1 binds to the...protein while others can inhibit CRISPR enzymes from multiple species of bacteria.  A note on anti-CRISPR nomenclature... of discovery (Bondy-Denomy et al., 2018). For example, AcrIIA4 mentioned above indicates that it’s the...activity one of three ways: Inhibit DNA binding Example:  AcrIIA4 blocks Cas9’s interaction with the PAM...site (Harrington et al., 2017). Inhibit crRNA loadingExample: AcrIIC2 interactions with Cas9 interferes ...and spatial control of CRISPR-Cas activity. For example, fusing AcrA4II to the photo-sensitive LOV2 domain...

  3. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    ...predecessors. Changes were implemented and I had to adapt accordingly. For example, the quarterly and annual...annual NPA meeting to learn more. Having met many people at that conference, I later became involved with...volunteered for various tasks as things came up. For example, I updated some sections of the NPA’s website. ...volunteering, you can gain experience working with other people mostly online and on monthly phone calls. This ...distance communication and learning to work with people despite the distance improved my verbal and written...understand how to work with many different types of people both above and below you. During my time at the...NPA. You must accommodate many different types of people in order to harness their strengths and work effectively...

  4. Experimenting in an Open Source lab: from CRISPR to Cats

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    Blog Post
    ...are often thought of as simple organisms , they have many levels of complexity, especially when trying...about it, the more I realized that it cuts across multiple disciplines, which really epitomizes what the ...learning how to grow and care for the worms. As an example of one of the problems we work collaboratively ...We attempt to deconstruct and understand this complexity using existing C. elegans mutants. For instance...data is freely available, and the collection of samples and sequencing is being organized and funded by...allows the project to be open to a wider group of people. This is a perfect starter project for our computer...Kittybiome project gives them access to rich and complex microbiome data for analysis. And, well, cats.....

  5. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    ...things started. Give a couple of audience members a chance to participate. For example, at a recent event ... Reddit AMA Perhaps we should start with some examples of a bad panel discussion? You know, the one where...interesting stories that will make for a fun discussion. People are much better at remembering these things on ... is tough for women in skirts, especially short people – which I am, so I hate stools). I like it when... with an unexpectedly great turnout). This gets people to sit close to the front so the interactions are...interesting discussion. The audience knows these people and bios were likely part of the publicity. Good...science or expertise by listing some and asking people to raise hands in response. I am always ready with...

  6. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...news is there’s a few simple ways to deal with this issue. One way is to grow multiple bacterial cultures...helps me figure out if I have enough plasmid to complete my experiment. AAV is usually the limiting reagent...concentrate the purified virus. This process can be completed in one long day, or the virus can be stored at...aliquots to use for qPCR titering. This helps avoid multiple freeze/thaws of your larger aliquots of virus....particles that are present in an AAV prep. AAV samples are first DNaseI digested to remove any residual...technology or a TaqMan primer/probe set can be used and samples are quantified by comparing to an AAV standard ... plasmid. This whole process takes ~3 hours to complete: 1 hours hands-on time and 2 hours for the qPCR...

  7. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...co-expression of multiple unique genes, you can start with a plasmid that has multiple cloning sites flanking...Co-expression of multiple genes is valuable in many experimental settings. To achieve this, scientists...co-transfection of two or more plasmids, the use of multiple or bidirectional promoters, or the creation of... are useful almost anytime you want to express multiple genes in the same cell. Although it is possible...to drive co-expression by using a plasmid with multiple, individual expression cassettes, having the genes...viruses to allow for the efficient translation of multiple genes from a single transcript. The two strategies...20 amino acids) and produce equimolar levels of mulitple genes from the same mRNA. The term "self-cleaving...

  8. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    ...in protein complex studies where tagging of multiple protein products is desired, as multiple selection...cerevisiae and S. pombe. The protocol is surprisingly simple, requiring only a PCR product containing the modifying...for easy capture and detection of proteins and complexes, without the artifacts sometimes associated with...Formosa, at the University of Utah, has built a complete collection of yeast tagging modules with each ... Dr. Formosa has deposited six plasmids with a multiple cloning site in place of the epitope tag for creation...ideal for tandem affinity purification of protein complexes. John Pringle and Jürg Bähler have deposited a... S. cerevisiae, with the additional benefit of multiple selection markers for combining modifications ...

  9. MXS Chaining

    Type
    Blog Post
    ...many biological fields. In synthetic biology for example, high-throughput cloning can be used to combine...powerful imaging technologies, the combination of multiple fluorescent protein sequences to develop diverse...one such technique and has been used to produce complex fluorescent reporter constructs. These fluorescent... variety of other clever applications. In one example, modular constructs generated with MXS-chaining...Tubulin Repeatable, directional cloning   The principle behind MXS-chaining is repeatable, restriction-enzyme...individual plasmids and are each flanked by the same multiple cloning site (MCS). Modules are assembled one ...being directional, meaning that (in the previous example) one can control whether the third module is added...

  10. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ... affected by gene activation rather than depletion. Multiple strategies have been developed to recruit... stranded DNA breaks that commonly result in a complete loss of function while nuclease-deactivated Cas9...and assessed the libraries performance via the depletion of a gold standard set of genes (1,580 essential...effective than other popular CRISPR libraries in the depletion of essential genes. In the subsampling analysis...have several limitations. CRISPRko results in a complete loss of function meaning that if an essential ...libraries utilize an engineered Cas9, which contains multiple point mutations in the nuclease domain. These ...domain such as VP16 to dCas9 or the recruitment of multiple copies of VP16 termed the “Sun Tag” method (Tanenbaum...

  11. What's New in CRISPR - March 2020

    Type
    Blog Post
    ...downregulate multiple genes at once. Read the paper in mBio Find the CRISPRi plasmids A multiplexed gRNA piggyBac...Base editing CRISPRi in Caulobacter crescentus Multiplexed gRNA for human pluripotent stem cells CRISPR-... Blue light renders the NLS accessible and the complex enters the nucleus where dCas9-Mxi1 can repress.... In the dark, the NLS is inaccessible and the complex remains in the cytoplasm. Read the paper in Cellular...differentiation, but it takes several weeks to delete or deplete individual genes. Michael Laub’s lab developed ... deliver uniform and sustained expression of multiplexed gRNAs in human pluripotent stem cells. Previously... construct and the fluorescently-labeled gRNA multiplex. Read the paper in Scientific Reports Find the...

  12. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    ...stimuli. One way they do so is through G protein-coupled receptors (GPCRs), receptors that play many roles...GPCRs (Takeda S et al., 2002) and many downstream complexes involved, it can be difficult for scientists to...in sight (Olsen RHJ et al., 2020). A transducer complex consists of a trimer of three subunits: alpha, ...collectively known as a heterotrimeric G protein. This complex converts a signal received by a GPCR on the cell...is being able to identify specific transducer complexes that are activated in response to a ligand binding...inactive, these subunits are found in a trimeric complex bound to the intracellular surface of the GPCR....Following activation in response to the ligand, the complex will disassociate, resulting in a decreased GFP2...

  13. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    ...approaches that may come up during discussion.  One example of agility at work is our process for working on...instructions on how to demonstrate the feature. For example, we recently reduced the number of steps involved...stories in that sprint relate to the theme. For example, we could have a shipping sprint and all the stories...get different perspectives on how things are implemented and from that, develop better software.  Maintenance...how you combine it with other ingredients.  For example, we use a third party library to handle label printing... compatible with the updated library. A recent example is updating our software from Python 2.7 to Python...analogy. Human languages evolve over time, for example from Ancient Greek to Modern Greek. Books written...

  14. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    ...there are some genetic factors involved. For example, multiple studies show that certain mutations of the...is a terrible disease that is, unfortunately, completely preventable. Mainly caused by exposure to asbestos...mesothelioma instead rely upon private donations. Many people think that because mesothelioma is so rare, there... researchers don’t entirely understand why some people develop the disease while others never do, but ...slow the growth of tumors. More often than not, multiple techniques are used as part of a treatment plan...positive potential for treating mesothelioma. For example, patients who have peritoneal mesothelioma – that... be used to distinguish between three groups of people:  Those who have mesothelioma Those who have been...

  15. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...fields in order to provide useful solutions to complex challenges. As a cross-disciplinary researcher,...Many of the challenges in science today are too complex for any single discipline to solve them on their...interdisciplinarity index of research for single or multiple disciplines – you can select the disciplines and...few years, I decided to look at what online tools people generally like to use to share ideas – with the... in terms of communicating ideas. In 2012, for example, 3 billion hours of video were watched each month... disciplines leading the way, the future of complementing published research with video overviews was ...networking, visual appeal, instant sharing across multiple other sites), I created a new platform for sharing...

  16. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ...field. Fundamental principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores...attached to self-labeling proteins. Some common examples of these self-labeling proteins would be SNAP-...of protein substrates/peptides and antibodies. Example applications of this include coupling to an antibody...specific receptors is key, especially for G-protein-coupled receptors (GPCRs). Mutations in receptor sites ...photons absorbed over the photons emitted. For example, DAPI is much dimmer than Alexa Fluor 488, and ...signal and facilitate clear differentiation between multiple dyes during analysis. Image resolution is impacted...microscopy techniques, such as Stimulated Emission Depletion (STED), and Stochastic Optical Reconstruction ...

  17. Science Careers: Unruly Interests Feed Many Paths

    Type
    Blog Post
    ...career Developmental biology caught me early on: a simple egg is packed with information. I am fascinated...individual diversity to get from gamete to a visibly complex - and unique - adult. My initial research directions...teacher was looking for an outlet. I started asking people: why do you do what you do? How do you like it?...it? how did you get there? Many of those people I met through AWIS (Association for Women in Science); ...visiting seminar speakers. My eye was caught by a couple of unusual job openings, although they seemed too...applied research. I focus in general terms on a couple of very broad topic domains, but each day brings...and surprisingly diverse manuscripts. I work with people, nurturing networks, as much as with technical ...

  18. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    ...adopting simple note-taking tools, others using excel sheets or organizing folders in complex ways, there...lengths to keep accurate records, but given the complexities of modern research, it’s often not enough. A...if they are to switch from paper to digital for example): “… 99% of respondents indicated that ease of ...developed the SciNote Electronic Lab Notebook. Implementing an electronic lab notebook starts with defining...property across various software providers used to supplement their paper notebooks. Instead, all of the important... you’ll be able to do it when needed. How many people from your lab would use the ELN i.e. do you need... facilitate this, some ELNs allow you to invite people and assign them roles and permission that only ...

  19. Transferable Skills Guide: Time Management

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    Blog Post
    ...SciComm skills and I had to coordinate with other people for a variety of projects. I was therefore forced...its utility) How much time it will take you to complete the task When picking which tasks to actually...accomplished quickly and things that will take longer. Completing the quick and useful tasks will swiftly give ... accomplishments will likely energize you. For example, let's say you’re doing a multi-day experiment ...experiment. The first way, it will take you 3 days to complete the entire experiment, but you’ll have to spend...experiment. The second way, it will take you 5 days to complete the experiment, but you’ll only spend half of ...remember things better when I write them down. Some people also find it useful to write out their to-do lists...

  20. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...proxy for dimerization (Shaner et al., 2008 - supplementary figure C). However, these assays are qualitative...fluorescent protein. These were either (i) coded by the multiple cloning site and therefore consist of random amino...protein is cleaved by proteases. Nowadays, with ample structural information available, it’s clear that...protein of interest (POI). To indicate the sequence, people often use terminology like “C-terminal fusion,”...#85045). We often use restriction enzymes in the multiple cloning site for generating fusions. For the POI-fluorescent... acids (PVAT, when we use the AgeI site in the multiple cloning site) is sufficient. For the fluorescent... better preserves the function of the POI. For example, the APT1-mVenus fusion localizes at the Golgi,...
Showing: 281 - 300 of 1009 results