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We narrowed to 850 results for: Met

Showing: 261 - 280 of 850 results

  1. Evolution of Lab Techniques

    Type
    Blog Post
    ...fluorescently labeled probes. Colorimetric and luminescence based detection methods are also commonly used. ...with fluorescently labeled ones allowing for fluorometric-based detection. This increased sequencing speed...sequencing machines (9). Today, similar technologies, sometimes referred to as sequence DNA as it synthesized ...fluorescently labeled nucleotides are incorporated, a method called pyrosequencing. The drive for faster, higher...typically rely on Sanger sequencing but instead use methods like pyrosequencing that are parallelized to sequence...of sequencing should bring efficient, low cost methods for sequencing large amounts of DNA quickly and...

  2. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...microbial opsins. Nature Methods, 9(2), 159–172. https://doi.org/10.1038/nmeth.1808 References Abreu, N...microbial opsins. Nature Methods, 9(2), 159–172. https://doi.org/10.1038/nmeth.1808 Nagel, G., Szellas,...multiple intra-cellular signaling cascades. Optical methods for controlling GPCR activation have been developed... than is achieved by traditional pharmacologic methods. A number of strategies have been developed for...mechanisms that are often cell-type specific. If it is something you’re considering for your experiment, note that...

  3. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...proteins from gels to diazobenzyloxymethyl-paper and detection with antisera: a method for studying antibody...western blot, or western, is one of the simplest methods to detect the presence or absence of a protein ...proteins typically do not transfer as well using this method.  Finally, commercially available dry blotting ...antibody, consider reviewing the materials and methods sections of those publications.  Finally, always...quantification of a protein, consider using an alternative method such as an enzyme-linked immunosorbent assay (ELISA...modifications such as phosphorylation, acetylation, methylation and ubiquitination. For example, to test if a...

  4. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ...gene, Mettl7a1, whose expression helped coax cells towards successful iEP reprogramming. MettI7a1 encodes...encodes an uncharacterized putative methyltransferase. Adding Mettl7a1 to the standard Foxa1-Hnf4α retroviral...CellTag CellTagging is a lentiviral based tagging method that labels cells with heritable barcodes. Barcodes...reprogramming and direct reprogramming Explore different methods for delivery of reprogramming factors for generating...

  5. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    ...the trick is to utilize a second immobilization method. One possibility is covalent modification of the...a similar function as GST in that it provides a method to express a foreign protein in bacteria and subsequently...determine binding kinetics using a multitude of methods including surface plasmon resonance (Biacore, GE... GE Life Sciences) or bio-layer interferometry (Octet, Pall ForteBio). Tips for using pMal-T-Avi-His/BirA...commercially available. Factor Xa loves calcium, but metal-chelate matrices, not so much. Maybe a calcium-independent...that have been biotinylated using less efficient methods. Many thanks to our guest blogger Jon Backstrom...

  6. Antibodies 101: Validation

    Type
    Blog Post
    ...validation of antibodies. Nat Methods 13:823–827. https://doi.org/10.1038/nmeth.3995  ...antibody-dependent approach. Antibody-independent methods may include analysis of transcriptomic or proteomic... between the levels of target detected in both methods. These approaches have relatively low sensitivity...be concerned that your antibody is binding to something unexpected. This approach has high specificity...epitopes of this protein.   You perform flow cytometry using both antibodies, as well as a third unrelated...sample using the antibody and then uses mass spectrometry to identify the captured proteins. If the antibody...

  7. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    ...launch a product? Make an informed decision about something? Fix a problem? Test a hypothesis? Whether you...to make progress towards open-ended goals whose metrics for success are defined by ongoing mission fulfillment...The benefits of a team So why is it helpful, or sometimes even necessary, to build a team in the first place...diverse group of people to provide that base. Sometimes there’s just too much work If you are one of those... frustrating than being on a leaderless team. Sometimes the leader just has to be the person to say “OK... guys, we need to have a meeting about this.” Sometimes the common roles of a leader can be successfully...expertise your current team members don’t have - sometimes you just don’t know who you need until the team...

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

    Type
    Blog Post
    ...Watch out for methylation issues. Enzymes like XbaI and ClaI are sensitive to methylation and their activity...sequence of all the plasmids we distribute. This method is time-intensive, so we recommend a variety of...sequence. Once you have purified plasmid DNA, this method can be done right in your lab in less than a day...will need to purify your DNA from a dcm or dam methylation-deficient bacterial strain such as JM110 or INV110...lanes look like your uncut lane then there is something wrong! Quantify your DNA. Loading too much DNA...

  9. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...development of multiple virus packaging methods. Across these methods, the following components are needed...Packaging using 293T cells This method is very similar to lentiviral packaging methods. The three components described...Packaging using helper-free packaging cell lines This method utilizes a packaging cell line that stably expresses... these genes in trans by specific plasmids. This method, therefore, reduces the number of plasmids that...

  10. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    ...mutated methionine was not enough to prevent alternative initiation, so they mutated the methionines at position...without any off target effects. However, these methods can be labor intensive and quite time consuming... will focus on two fluorescent CRISPR reporter methods, SRIRACCHA and GEMCherry2. We will cover single...start site initiation, the lab removed the native methionine start site. This first iteration was named GEmCherry1...for your CRISPR mutant of interest, give these methods a try.  References Højland Knudsen C, Ásgrímsdóttir...

  11. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    ...eukaryotic cell through non-viral methods including both chemical and physical methods in the lab. Chemicals like... the lab. Let’s take a look at these different methods of DNA insertion. Transformation Transformation...conjugation Conjugation was the first extensively studied method of gene transfer and was discovered in 1946 by ...Kostman.   Transfection Unlike the last three methods which can be used in prokaryotes, transfection ...the negatively charged cell membrane. Physical methods such as electroporation or microinjection actually...

  12. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...experiments. MET15 L-methionine and overproduces hydrosulfide ions yes - Grow with methylmercury. S. cerevisiae...to high copy numbers. This creates a potential metabolic burden on the yeast cells. Some phenotypes may...tried varied approaches to combat these issues. One method to reduce the amount of marker gene expression ... utilizing the more traditional drug selection methods feasible in yeast as a complement or alternative...

  13. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...swamped out by a sea of uncoupled donors. The best method for targeting a fluorescent donor to your protein...tRNA synthetases is required and thus far, this method has proven most effective only in a few cell types...This is not an extensive list Acceptor labeling: Sometimes it's okay to be wrong In contrast to FRET donors... (which was originally developed to treat heavy metal intoxication in chemical warfare [Vilensky & Redman... of your FRET assay (Figure 2). These labeling methods are freely interchangeable and new uses for FRET... structure using a novel site-specific labeling method." PloS one 4.10 (2009): e7338. PubMed PMID: 19823671...dynamics in living cells with FRET reporters." Methods 40.3 (2006): 279-286. PubMed PMID: 16908183. 10...

  14. The time and cost required to make a plasmid

    Type
    Blog Post
    ...What about all the reagents you need to order? Sometimes, it feels like an eternity of cloning, waiting...costs of producing plasmids with high-throughput methods that save on reagents, consumables, and time. We...first try (or any response at all). And while this method is free for you, and many labs are quite happy ...strategies beyond the traditional PCR/digest/ligate methods and picking the right one can save you time if ...speed or money if budget is a concern. The faster methods, like Gibson assembly, are typically more expensive... 5 factors to help you choose the right cloning method Find blog posts about scientific sharing here Download...

  15. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    ... a visual aid instead of describing something verbally. Sometimes a group was just really tired and needed...as it does to read an 11-page Cell paper - and sometimes, it might actually take me less time to read the...concepts are easier to grasp than others. But sometimes we’re unaware that we are leaning on an unspoken...way I thought about engaging with learners. If something I was communicating didn’t click, instead of blaming...put their phones away), or maybe I was assuming something was intuitive that actually wasn't. Maybe I needed...

  16. Oh, The Places You Can Go: Careers in Science Communication - Writing for a Research Institute

    Type
    Blog Post
    ...’s science communication journey started with something unexpected: termites. “I was really interested... work for one client and then switch to doing something else. You’re never really integrated and part ...pulled in all of these different directions is sometimes at odds with the space needed to be creative. ...a lot of time to let you mind process things. Sometimes finding that uninterrupted time to be creative...some articles are just harder to write. “There’s something about being a writer that can be a very painful...these problems through connections and that’s something that good storytellers in science can do.” Susan...

  17. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ...you’re probably aware that there are several methodologies currently available for approaching it. These... traditional restriction enzyme/ligase-mediated method, the more recently developed Gibson Assembly Cloning...cloning technologies, as well as several others. Each method is unique and relies on specific components that...components is essential for choosing the correct cloning method for your own experiments, and here we will focus... a unique gene that makes the popular GatewayTM method possible: ccdB. But what is ccdB, what role does... to ensure that you choose the most appropriate method for your own experiments. Please check out Addgene...

  18. Working with Nuclear Receptors

    Type
    Blog Post
    ...largest family of transcription factors found in metazoans. The superfamily is comprised of 48 human and ...a broad range of reproductive, developmental, metabolic, and immune response programs. NRs also encompass...gallstone disease, senescent illnesses, and the metabolic syndrome. Coregulators Coregulators constitute... regulatory regime (such as genes involved in metabolism, growth or cytokine action). Primary or core ...and CBP histone acetyltransferases or CARM1 methyltransferase. This attendant signaling feature afforded...adipogenesis and macrophage activation. Our mass spectrometry-affinity purification proteomic profiling of...

  19. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ...interrogate neuronal networks. A major drawback of this method is the stray excitation of neighboring neurons ...material for the bioproduction of therapeutic methylxanthine drugs like theophylline. To facilitate future...enables the degradation of caffeine (1,3,7-trimethylxanthine) and its conversion into xanthine. The majority...constitutive promoter (BBa_J23100) and expresses N-demethylases and other required enzymes, including a glutathione...rescued by the xanthine produced via complete demethylation of caffeine. For example, the Barrick lab was...Find the FLINC biosensors here. Mo et al. Nat Methods. 2017. PubMed PMID: 28288122. New fluorescent...

  20. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ... to some of your potential options and cloning methods, but please scroll down for plasmids suitable for...inserted using Gibson Assembly or SLIC cloning methods. A BsaI-based E. coli multiplexing plasmid is available...multiplexed gRNAs with Golden Gate and Gibson assembly methods If you want to scale up the number of gRNAs in ...your plasmid, you’ll need to use some assembly methods such as Golden Gate or Gibson assembly. Golden ...containing destination vector. Note: depending on which method you use, the procedure will vary slightly. ...Fok1-dCas9 (dimeric nuclease). Gateway assembly method Frew Lab Multiple Lentiviral Expression Systems...Plasmids: The CRMAGE system is a fast, multiplexable method that combines CRISPR and recombineering-based MAGE...
Showing: 261 - 280 of 850 results