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We narrowed to 712 results for: Met

Showing: 181 - 200 of 712 results

  1. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    Published
    Nov. 28, 2017, 2 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Aug. 28, 2014, 3:34 p.m.
    ...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...

  3. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    Published
    Aug. 6, 2024, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    June 25, 2019, 12:54 p.m.
    ...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...

  5. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    Published
    May 26, 2020, 1:15 p.m.
    ...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...

  6. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    Published
    Feb. 25, 2014, 7:11 p.m.
    ...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...

  7. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Feb. 2, 2018, 3:20 p.m.
    ...’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...

  9. The time and cost required to make a plasmid

    Type
    Blog Post
    Published
    March 23, 2023, 1:15 p.m.
    ...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...

  10. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    Published
    June 16, 2022, 1:15 p.m.
    ... 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...

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

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ...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...

  12. Working with Nuclear Receptors

    Type
    Blog Post
    Published
    July 15, 2014, 4:03 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ...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...

  14. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ... 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...

  15. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    Published
    Sept. 17, 2024, 1:15 p.m.
    ...ice, whichever is more practical. If your lysing method is not resulting in your protein being available...need such precise measurements, you can use a colorimetric ladder, visible to the naked eye; a fluorescent... of protein, make sure your reversible staining method of choice is appropriately sensitive. If using ...membrane, you can dry the membrane, wet it with 20% methanol, and place over a light box to visualize proteins...that the p membrane pores are too large.  Transfer method   Wet transfers are more efficient, so if you’re...re-run the western.  Remember that chemilumescent methods, like horseradish peroxidase protein (HRP), have...the bubbles in the membrane sandwich Ensure methanol concentration in transfer buffer is < 20% ...

  16. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ...You have to weigh the pros and cons of various methods and choose what is best for you — and the mice....potential pain or discomfort of the mice, and restraint method. Many different substances can be injected, including...on the back. Depending on the site and restraint method, subcutaneous injections take some practice to ...injections is studying inflammatory responses, as this method is useful for analyzing localized events. Intradermal...the throat. The feeding tube can be flexible or metal, depending on researcher preferences. It sounds ...injection routes in mice! When considering what method you’d like to use, remember that the final decision...

  17. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    Published
    Nov. 5, 2024, 8:32 p.m.
    ... production or looking to improve your current methods, understanding the differences can help you make...will depend on your specific virus, so this is something you may have to optimize for your specific application...the virus type. AAV is typically purified using methods like ultracentrifugation, while lentivirus supernatants...up to 38 kb! Generally, insect cell production methods produce lower titers of virus than mammalian systems...systems. This is why many insect production methods add an additional viral passage. This involves taking...taking your first round of virus, called P1 (or sometimes P0), infecting new Sf9 cells, and getting an amplified...

  18. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ...of interest and examine them using biochemical methods thus removing the context of the cell. However,...important to choose the appropriate quantification methods and tools. It is usually not enough to examine ...review of quantitative colocalization analysis methods and useful image analysis software tools. Pitfalls...Pitfalls, limitations, and specialized colocalization methods Fluorescent proteins are very useful tools, but...further investigation. One fluorescent microscopy method commonly employed for analyzing interactions between...fluorescent protein for cellular imaging." Nature Methods 14.1 (2017): 53-56. PubMed PMID: 27869816. 6. Media...

  19. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    Published
    Sept. 28, 2017, 2:01 p.m.
    ...AAVs for neuroscience research is the lack of a method for genetically manipulating neurons throughout... Plasmids Here! Their Solution: Use the CREATE method to engineer two new IV injectable AAV capsids, ...for studying the CNS and PNS! What is the CREATE method?  CREATE stands for Cre Recombinase-based AAV Targeted...axons and dendrites extend to. The keys to this method are 1) high levels of color diversity so that cells...system is less cumbersome than brainbow, another method for multi-color labeling of neurons, because it...capsids (figure 1) were engineered with the CREATE method (figure 2) to allow for efficient IV delivery of...

  20. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    Published
    Jan. 16, 2020, 2:18 p.m.
    ...are in the process of transitioning our titering method from qPCR to ddPCR. Droplet digital PCR (ddPCR)...your PCR For a good starting place on what PCR parameters to use, see Lock’s seminal paper on AAV titration...between positive and negative droplets with these parameters, here are a few things you can try: Increase...in faecal samples." Journal of microbiological methods 148 (2018): 64-73. PubMed PMID: 29548643. Lock,...titers by droplet digital PCR." Human gene therapy methods 25.2 (2013): 115-125. PubMed PMID: 24328707. PubMed...Kristina, et al. "A systematic investigation of parameters influencing droplet rain in the Listeria monocytogenes...
Showing: 181 - 200 of 712 results