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  1. Save Time with Transient Plant Leaf Transformations

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    Blog Post
    ... lot of time. Don't re-invent the wheel. Test different leaf ages Usually younger leaves work better. ...luciferases to more easily quantify the effect of the different treatments on transformation success. Some reporter... all plasmids from the Lee-Parsons lab. Test different leaf infiltration methods Co-cultivation (Li ...in the plant but not from Agrobacterium. Test different Agrobacterium strains We used the disarmed Agrobacterium...allows you to quickly assemble vectors and test different ideas. We are a huge fan of the Golden Gate based...starting his work in the Lee-Parsons Lab.     References D'Aoust, Marc-André, et al. "Transient expression...
  2. Rett Syndrome: A History of Research and Therapeutic Outlooks

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    Blog Post
    ...Austrian physician, Andreas Rett, who tried to raise awareness and published his observations in a German medical...hope that a similar reversal may be possible for children and adults with Rett Syndrome.  Since these discoveries...discoveries, scientists have studied how different parts of MECP2 influence Rett Syndrome. Zoghbi’s lab...varying degrees determine the clinical severity of different MECP2 mutations. More recently, the Bird lab found..., Rett databases, and other helpful tools.  References: Amir RE, Van den Veyver IB, Wan M, Tran CQ, ...Extends the Survival of MeCP2-Null Mice without Apparent Toxicity after Intracisternal Delivery. Molecular...
  3. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

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    Blog Post
    ...using FACS which could limit their use across different cell types and systems. To introduce flexibility... GO works by affecting protein translation of different reporter proteins. Protein translation requires...used GO to induce the translation of an array of different reporters including mScarlet-I, luciferase, antibiotic...proteins, antibiotic resistance, and luciferase. References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran.../10.1093/nar/gkaa124 Martin AS, Salamango DJ, Serebrenik AA, Shaban NM, Brown WL, Harris RS (2019) A panel...s41598-018-36739-9 St. Martin A, Salamango D, Serebrenik A, Shaban N, Brown WL, Donati F, Munagala U, ...
  4. CRISPR-based Genome Editing Systems For Editing in E. coli

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    Blog Post
    ...CRISPR-associated transposition.     References and Resources References: Jiang Y, Chen B, Duan C, Sun B, ... E. coli host, and the addition of a sacB gene renders it beneficial for plasmid clearance. It is more...concatenations into crRNA arrays targeting multiple different sites in E. coli. pCutamp cures the plasmids by...MUCICAT toolkit includes versions of plasmids with different induction systems, such as PT7, Prha, Ptet, and...Capability to construct a strain library with different gene dosages to facilitate swift optimization ...
  5. New CRISPR Tools: Cas7-11 and PASTE

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    Blog Post
    ...forward for what the field can do.    References and Resources References  Özcan A, Krajeski R, Ioannidi E,...development of tools that exploit and expand our current knowledge. The Abudayyeh-Gootenberg lab (also called... was sent to the AbuGoot lab for testing, they weren’t sure what they would find… but it was safe to say...fibrosis, which can be caused by thousands of different mutations, all found in the same gene. If we only...the number of potential applications. But they weren’t done engineering quite yet – surely, they thought...
  6. CRISPR Activation: A Practical Guide

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    Blog Post
    ...example, CRISPR activation has been used to differentiate induced pluripotent stem cells (Chavez et al...neuronal cells by activating genes that cause differentiation and to reverse HIV viral latency (Bialek et...already be hitting an upper bound of activation that current Cas9 systems cannot help you pass. In addition,...directed towards that gene as there can be a large difference in guide potency. Thus, in cases where users ...Chavez et al., 2016). To learn more about the different types of Cas9 activators, check out our blog post...DNA sequence or regulating gene expression.     References Behan FM, Iorio F, Picco G, Gonçalves E, Beaver...
  7. Tips for CRISPR Gene Editing in Mice

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    Blog Post
    ... to determine the pros and cons of using many different types of gRNA design software. When you are happy...yourself through whatever DNA supply company you currently use. You can also find many previously validated...inserted or deleted. Each lab will use a slightly different method to genotype their pups. Taking a toe, a...weeks old and use the ear tag method (creating different patterns of ear holes and using that material ...latest and greatest in scientific content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target...plasmid into zygotes." Development, growth & differentiation 56.1 (2014): 122-129. PubMed PMID: 24372541...
  8. Replacing Paper: Tips for Choosing an Electronic lab Notebook

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    Blog Post
    ...labs. While many labs approach the situation differently, some adopting simple note-taking tools, others...functionalities with experiment records. Many different types of ELNs are available on the market today...available here. Our questions focused on researchers’ current practices, their opinions towards ELNs, and their...desired functionalities i.e. main priorities of different ELN features, ranging from whether respondents...cases this applies to universities. How are you currently exchanging information with your team? If you ... allows you to assign specific permissions to different lab members to avoid any issues later on. Do you...
  9. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

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    Blog Post
    ... iPSCs (or correct them back to wild type), differentiate them into somatic cells affected by the disease...iPSC clone picking, a 100-fold decrease is the difference between picking 100 versus 10,000 clones! We ... experiments with blocking mutations, we still weren’t entirely happy with our editing outcomes and in...Dominik Paquet and Dylan Kwart Dominik Paquet is currently a Professor for Neurobiology at the Institute ...Tessier-Lavigne’s lab at the Rockefeller University. He currently works to understand pathological mechanisms of...developing novel iPSC-based model systems.     References 1. Cong, Le, et al. "Multiplex genome engineering...
  10. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

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    Blog Post
    ...tagged protein because of its size. This type of interference is known as steric hindrance. For instance, ...OSER whorls. This assay is quantitative and is currently the standard for assessing oligomerization in ...fluorescent protein. I prefer to describe the different parts of the fusion from N- to C-terminus. For...advisable to generate multiple constructs with different insertions, since it is difficult to predict which...can follow him on twitter: @joachimgoedhart.   References Adjobo-Hermans, Merel JW, et al. "Real-time visualization...291.5512 (2001): 2408-2411. PubMed PMID: 11264536. Klarenbeek, Jeffrey, et al. "Fourth-generation epac-based...
  11. Plasmids 101: Biotinylation

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    Blog Post
    ...solubility, and thus different reagents are useful for biotinylation in different microenvironments. There...used today in molecular biology for an array of different techniques and protocols. In this post we will...determined to be due to a deficiency in Vitamin H later renamed Vitamin B7/biotin. Egg white injury would plague...a solid substrate (Holden and Cremer, 2003). References 1. Chapman-Smith, A., & Cronan, J. E. “Molecular...post-translational modification of exceptional specificity.” Trends in Biochemical Sciences, 24.9 (1999b): 359–363....
  12. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ...sensors with different ITAM/SH2 reporter pairs allow multiplexed measurement of different RTKs. Although...Improving on the glutamate sensor iGluSnFR from Loren Looger’s lab, Kaspar Podgorski and colleagues have...response. They generated variants optimized for different applications (v82 has higher affinity for glutamate...By fusing two bright fluorescent proteins with different thermal quenching rates (mNeonGreen and tdTomato...sub-millisecond resolution and sub-degree temperature differences under a variety of imaging conditions (155 μs...
  13. Five Popular Model Organisms, Part 2

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    Blog Post
    ...similarities with humans. However, rats have some key differences in physiology, intelligence, and human correspondence... generate transgenic axolotls for an array of different studies. Elly Tanaka’s lab has constructed several...include understanding of allelic variation between different plant species and metabolic properties affecting... collections to help start your experiments! References: Blount, Z. D. (2015). The unexhausted potential...Zhang, C., & Xing, X.-H. (2018). Pooled CRISPR interference screening enables genome-scale functional genomics...Plasmids Collection Page Read our molecular biology reference Find plasmids for your research ...
  14. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

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    Blog Post
    ...scientists have developed and fine tuned many different ways to clone DNA fragments which have provided...with an enzyme master mix, which contains three different enzymes: an exonuclease, which chews back the... this doesn’t allow for mix and match of many different fragments. One way around this is to use a combination...with a G!) but actually work by significantly different methods. Golden Gate cloning does result in the...from one vector to another. There are lots of different ways to clone these days. Gibson and the other...biology, you should consider giving it a try! References: 1. Gibson DG, Young L, Chuang RY, Venter JC,...
  15. Keeping Up With CRISPR/Cas9

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    Blog Post
    ...available at Addgene CRISPR-Cas9 References, Protocols, and more References: Generation of Gene-Modified ...the monkey story hit the presses, another very different bit of CRISPR news was circulating. Scientists...how CRISPR/Cas9s actually work. Last month, the Lawrence Berkeley Lab's Jennifer Doudna and Eva Nogales...this structure may help us engineer around the current limitations of the Cas9 complex," said study author...
  16. Plasmids 101: Repressible Promoters

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    Blog Post
    ...by Hannah Dotson in Dec. 2022.  References and Resources References Gossen M, Bujard H. Tight control...In the presence of tetracycline (Tet), TetR preferentially binds Tet over the TetO elements, allowing ... One can therefore place GAL4 and GAL80 under different promoters and create sophisticated patterns of...wondering, “why is it necessary to have all 3 of the different binary systems above if they all behave very similarly...tet-on-and-tet-off-2nd-generation Mullick, A., Xu, Y., Warren, R. et al. The cumate gene-switch: a system for...
  17. PiggyBac-ing Through the Genome Editing Field

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    Blog Post
    ...line with the piggyBac™ system is that it has a preference for integration at regions of open chromatin,...rearrangement (such as when a stem cell initiates differentiation), transgene silencing may occur (as it would...cassette can be incorporated, and used during the differentiation process. Moreover, a splinkerette-PCR assay...technologies and strategies for their projects. References 1. L. E. Woodard and M. H. Wilson, “piggyBac-...piggyBac-ing models and new therapeutic strategies.,” Trends Biotechnol., vol. 33, no. 9, pp. 525–33, Sep. 2015...26299960. PubMed Central PMCID: PMC4559351. 8. A. G. Uren, H. Mikkers, J. Kool, L. van der Weyden, A. H. Lund...
  18. Science Communication for Everyone

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    Blog Post
    ...Toronto, created The Addictive Brain to spread awareness about how scientists utilize tax-payer money and... how our articles have helped them understand different topics of science, especially disease related ...terms like manipulations, anomaly, and so on, differently than scientists would. For example, “manipulate...blogger, Chinmaya Sandangi! Dr. Chinmaya Sadangi is currently a postdoc at the University of Toronto and founder... receive our science career guide Learn about different careers in science communication Resources on...
  19. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

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    Blog Post
    ...fragments that are very different.” Because the fragments produced will have different overhangs, they can...systems. “The cloning is all done in vitro; it’s no different in mouse, human or plants,” explained Jan Lohmann...Lohmann of Heidelberg University. “What is different is the plasmids you use to bring this into your target...available, making it easier for us to do our work.” References 1. Athanasios Lampropoulos et al. "GreenGate ...
  20. The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years

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    Blog Post
    ...5,000 different research institutions. This has given us a unique window into technology trends in the...by the influx of plasmids for reprogramming differentiated cells into pluripotent stem cells. Plasmids...HA-Ubiquitin-WT and HA-Ubiquitin, and we now have 19 different ubiquitin plasmids in the repository. 100,000 ... and modified by so many labs and for so many different purposes that we had to create a CRISPR page to...
Showing: 481 - 500 of 1013 results