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We narrowed to 850 results for: met

Showing: 241 - 260 of 850 results

  1. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...the “elegans” methods outlined in this post – particularly the use of co-selection methods – could greatly...templates in C. elegans. Methods 121–122:86–93 . https://doi.org/10.1016/j.ymeth.2017.03.023 Paix A, Wang...exhibit complete embryonic lethality at 25 ºC. This method resulted in efficiencies ranging from 11-100% of...selection allowed me to optimize a range of editing parameters: oligo repair templates with homology arms of...years:  Repair template design The Meyer lab meticulously characterized several aspects of editing. First...Cas9 Since the development of CRISPR/Cas9 editing methods, other Cas proteins have been adapted for use in...

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

    Type
    Blog Post
    ...O, Zhang F. Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047. 10.1038/nmeth.3047. PubMed PMID...add 3 microliters of 5 mM star anise?” This is something I was expecting when I set out to make my first...obtained through Addgene’s viral service and the methods sections of scientific articles (gasp). To my delight...the duplex and run it as an undigested control. Sometimes the PCR products can look smeared, so having an... not (Figure 4). 5. In theory you could use densitometric analysis to quantitate the intensity of the ...knockout a single allele in a human line, using methods based on homologous recombination (Rago et al.,...

  3. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    ...He has recently developed a third-generation intensiometric glutamate sensing fluorescent reporter (iGluSnFR3...biosensors or biosensors that detect new redox metabolites are vital for researchers to better understand...withdrawal before they occur.  First, she used a genetic method to tag neurons activated by opioid withdrawal with... opioid withdrawal, providing insight into new methods for treating patients in recovery from opioid use...is in progress, she has validated the dual virus method to study CR neurons which are depicted here. (A...biosensors based on their dynamic properties. Her method takes a pooled library of mutant sensors expressed...engineer many types of biosensors.   This screening method allowed Tian to optimize a red‑shifted archaerhodopsin‑derived...

  4. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    ...Aveiro, Portugal. If cancer was a musician, then metastasize is touring. But you know what a touring musician...divided into 3 phases: initiation, progression, and metastasis. The microenvironment’s physiology is deeply ...the tumor. How do microenvironment enzymes and metabolic reprograming influence tumors development? Being... vice versa, let’s take a look at cancer cell metabolism. Cancer cells promote oxidative stress leading...altered fibroblasts and promote the development and metastasis of tumors through inciting inflammation. Speaking...of cells affect tumor progression, enzymes and metabolism reprogramming aren’t the whole story. It’s essential...Tumour exosome integrins determine organotropic metastasis." Nature 527.7578 (2015): 329. PubMed PMID: 26524530...

  5. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    ...of microbial opsins. Nature methods. 2012;9(2):159-72. doi: 10.1038/nmeth.1808. PMID: 22179551; PMCID:...run it? And, most importantly, what are the methodological and experimental limitations and considerations...considerations you may not find in the materials and methods sections of optogenetics papers. Optogenetics...Delivering your opsin  The standard opsin delivery method is local injection of an adeno-associated viral...may need to adjust coordinates based on your own method of identifying bregma but it is best to spend the...stimulation. Testing opsin expression  Regardless of the method of expression of your opsin gene, it is essential...dissection of neural circuitry function. Nature methods. 2011;8(9):745-52. PMID: 21985008; PMCID: PMC3191888...

  6. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    ...topic.  Meta Meta is a literature discovery platform developed by the Chan Zuckerberg Initiative. Meta is ...number of manuscripts that cite the background, methods, or results section is also displayed. Saving ...interest. You can also just follow feeds curated by Meta. Feeds automatically update as papers are published...number of times a paper is shared online, or its altmetric score, is also displayed in your feed. By clicking...whole.   Saving papers Papers are easily saved to a Meta library. The library will suggest additional papers...

  7. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...neuronal tracing methods Traditional tracing compounds either produce a colorimetric product or are fluorescent...technology, new methods have been developed to study neuronal connectivity. These methods are less neurotoxic...neuroscience methods such as electrophysiological recordings. The origins of these methods are naturally...inserted electrodes) may be applied, but these methods can have a negative impact on cellular health. ...versus Intradiaphragmatic Injection. J Neurosci Methods 241, 137–145. PubMed PMID: 25555356. PubMed Central...for single- and double-label studies. J Neurosci Methods 55, 65–78. PubMed PMID: 1381034. Watakabe, A., ...

  8. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...as the method is dependent on number of samples rather than number of cycles.  Choosing a method  As with...real-time PCR. J Microbiol Methods. 2006 Jun;65(3):476-87. doi: 10.1016/j.mimet.2005.09.007. Epub 2005 Oct...confidence in the results you need.  Whichever method you choose, resist the temptation to over interpret...Copy Number Variation with Droplet Digital PCR. Methods Mol Biol. 2018;1768:143-160. doi: 10.1007/978-1...quantitative real-time polymerase chain reaction methods for the determination of amplification of Plasmodium...

  9. Twelve Amazing Guest Blog Contributors!

    Type
    Blog Post
    ...shared his flow cytometry expertise in three posts. An Introduction to Gating in Flow Cytometry, Designing...shared her flow cytometry expertise in two posts. Flow Compensation and Flow Cytometry Controls Priyamvada... guide to understanding your flow data. Flow Cytometry Readouts Toj, a summer intern who shared their...validation expertise. Antibody Validation for Flow Cytometry Rob Hurt, whose expertise and suggestions were...

  10. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...scientists are no exception to this effect -- meticulously designed experiments in labs around the world...modification also eases the interpretation of the parameters we infer, since scientific productivity is better... model to give uncertainty estimates for each parameter, we added log-normal noise to each of the terms...gave us the maximum likelihood estimates of each parameter. We approximated the data likelihood with multivariate...dates impacted by each effect. The inferred parameters suggest that COVID-19 induced a 2x–5x reduction...the instability and uncertainty in estimating parameters from relatively sparse data from China. An important... uses applied machine learning and statistical methods for fundamental scientific discovery and high-impact...

  11. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...target metabolite creates an aptamer that will only bind to DFHBI in the presence of that metabolite (Fig... led by Jack Szostak and Larry Gold, developed methods for selecting and amplifying RNA sequences that...including proteins, peptides, amino acids, drugs, metal ions, and cells (even pathogenic bacteria). That...to incorporate their target molecule, such as a metal ion, or fold to incorporate themselves into a larger...difficult-to-study RNA-modifying enzymes, such as the RNA demethylases FTO and ALKBH5. Their ease of production have...2. Baker, Monya. “RNA imaging in situ”. Nature Methods 9, 787–790 (2012). 3. Filonov, Gregory S., et al...

  12. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...-mutant rabies virus. Nat Methods 4:47–49 . https://doi.org/10.1038/nmeth999 Wickersham IR, Lyon DC, Barnard...by nonspecific complement mechanisms in serum. Sometimes this is due to the cells that the viral vectors...Scientists have been using similar pseudotyping methods with other viruses in a BSL-2 lab instead of a ...the upregulation of LDLR expression using other methods.  In terms of preparation, VSV-G is stable and ...given brain region (Mazarakis et al., 2001).  This method can also allow scientists to trace a neuron’s projections...Vectors: One Vector, Many Guises. Human Gene Therapy Methods 28:291–301 . https://doi.org/10.1089/hgtb.2017.084...

  13. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    ...genders, educational, and professional backgrounds. Sometimes this can lead to wildly different ideas of how...our work.  Agile at Addgene We follow an agile methodology of software development called Scrum. The Scrum...during checkout. Demo details for this would be something along the lines of showing that the checkout takes... decide how to best approach it in our code.  Sometimes while thinking through these steps, we discover...more in-depth knowledge of one area, and when something urgent comes up, that person is the first in line...In some parts of our code, we might be calling something a sample and in other places, we might be calling...clean, reusable code when working on a feature, sometimes we have to take short cuts or add a band-aid to...

  14. Lab to Office Culture Shock

    Type
    Blog Post
    ...settings: The sense of being the first to discover something. The thrill of having an experiment yield clear...if I was asked that question today, because I sometimes wish for a stretch of time that doesn’t involve...mentally challenging than lab work at times.” Sometimes a desk job is defined as sitting down at a computer... between tubes, look through microscopes, and sometimes you even wear a lab coat! These are the things...is that I’ve never been bored. There is always something to do and there is no thumb twiddling. But you...time to come to realize that meetings were not something that took me away from my work. The meetings often...share their knowledge with the community (and sometimes conduct their own independent research!). There...

  15. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...CRISPR-Cas9 genome engineering parameters via a library-on-library approach. Nat Methods 12:823–826 . https://doi.org...transcriptional programming. Nat Methods 12:326–328 . https://doi.org/10.1038/nmeth.3312 Chavez A, Tuttle M, ... in multiple species. Nat Methods 13:563–567 . https://doi.org/10.1038/nmeth.3871 Gilbert LA, Horlbeck.../doi.org/10.1038/nmeth.3473 Chavez A, Scheiman J, Vora S, Pruitt BW, Tuttle M, P R Iyer E, Lin S, Kiani...

  16. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...isolated from a methylation competent bacterial strain (e.g. DH5α which is dam+). Methylation deficient bacterial...and Daniel R. Marshak. "Dimethyl sulfoxide-mediated primer Tm reduction: a method for analyzing the role...template. The parent template is removed using a methylation-dependent endonuclease (i.e. DpnI), and bacteria... of the primers. Because you will be using a methylation-dependent enzyme (DpnI) to eliminate the parent...is unique in that it cleaves only DNA that is methylated at the adenosine of the GATC recognition site...

  17. 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...

  18. 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...

  19. 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...

  20. Exploring Applications of the Bioluminescent HiBiT Tag

    Type
    Blog Post
    ...fluorescence imaging. Nature Methods, 14(7), 657–661. https://doi.org/10.1038/nmeth.4344  Peach, C. J., Kilpatrick... antibody-based detection or western blotting, methods that are low-throughput, semi-quantitative, and...substrate addition and a simple add-mix-measure method. NanoBiT® luciferase activity is compatible with... the endogenous locus or used other expression methods. Cracking the code of 'undruggable' targets Researchers...cells with pro-oncogenic traits, facilitating metastatic spread. The authors used NanoBiT assays to quantify...associated with endosomal localization of EGFR. This method offered kinetic, non-invasive monitoring of receptor...
Showing: 241 - 260 of 850 results