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Showing: 361 - 380 of 1082 results

  1. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...licensed distributor) allow you to quickly transfer a DNA sequence from the transposon vector to one of...encourage you to deposit your piggyBac™ transposon vectors with us to help us expand this useful resource....such as with signaling molecules or transcription factors, or when the gene is constitutively-active or a...edit of interest. With the larger piggyBac™ donor vector, it is possible to make the ITR and selection cassette...re-cutting. Transposase removal of the cassette later restores the target sequence. This allows researchers to...Transposase Read up on Genome Engineering with FLEx Vectors  Learn how Homology Directed Repair Is Used for...Genome Editing Resources on Addgene.org Find Vectors with the piggyBac™ Transposon Catch up on Everything...

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

    Type
    Blog Post
    ... DNA sequence called the Fertility factor (F-factor). The F-factor is found on an episome, a piece of ... information on viral vectors, including transduction download our Viral Vectors 101 eBook. Bacterial ...fragmented DNA released into the environment. Various factors promote natural transformation in different bacteria...bacterial or eukaryotic cells via a virus or viral vector. One example are bacteriophages that attach to ...scientists commonly use phagemids, a DNA cloning vector that contains both bacteriophage and plasmid properties...

  3. How Addgene Creates Content

    Type
    Blog Post
    ...entire mission is to enable that process. Our repository of plasmids, AAVs, antibodies, and data allows...technical skills, and experiences from thousands of laboratories that makes up our Help Center, blog, website... the blog originally started as a way to easily store and share in-depth answers to frequently asked questions...on a case-by-case basis, as many of our blog contributors primarily write in the areas in which they have...graphic designer upon request that the science communicator "just sketch a western blot out quickly." Final...and cited figures from papers or image sharing repositories. Then we format, do a final review to catch ...informative resources that answer questions our depositors and customers are asking. So how can you get ...

  4. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ...reporters require a chemical substrate (and sometimes cofactors) to be provided and are generally dimmer than ...Kinetics of photon generation Slower Faster Cofactors/substrates Required Not required Signal strength...engineered Peak emission (nm) Size (kDa) Substrate Cofactors Secreted North American firefly (Photinus pyralis...expressing luciferase Luciferase reporter assays for monitoring gene regulation Since firefly luciferase was ...expression analysis to investigate the effect of regulatory elements — such as promoters, enhancers, or untranslated...untranslated regions — on gene expression. The regulatory element is cloned upstream of the luciferase ...energy from an excited donor fluorophore to an acceptor fluorophore, which then emits it as light. Bioluminescence...

  5. Dos & Don'ts When Publishing a Scientific Manuscript

    Type
    Blog Post
    ...contributed by Maaike Pols, PhD, a Developmental Editor at F1000Research.  With many changes taking place...their funding should also be published. All these factors will influence your choice of journals to submit...communities. The ARRIVE guidelines for the reporting of laboratory animals during in vivo research and the CARE ... are two sets of guidelines frequently used by editors. Providing all this information makes your manuscript...who the reviewers are). If the reviewers and the editor decide that your manuscript needs to be revised...the conclusions of your paper. This will help the editor handling your manuscript decide on whether your... is appropriate for that journal. Argue with an editor or reviewer over their decision. If you do object...

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

    Type
    Blog Post
    ...proteins, such as human growth factor, vascular endothelial growth factor, and interleukin-6 (Witkiewicz... was contributed by Bárbara Pinho, a science communicator at the Portuguese science museum "Fábrica Centro...myofibroblasts, neuroendocrine, adipose, immune, inflammatory, angiogenic vascular, endothelial, and stromal... in increased expression of transforming growth factor-β signaling, possibly inducing the secretion of...seems to attack us by releasing mitogenic growth mediators and improving cancer cells proliferation.  How...enzymes and metabolism reprogramming aren’t the whole story. It’s essential for a tumor to spread and to resist... Pinho! Bárbara Pinho is currently a science communicator at the Portuguese science museum "Fábrica Centro...

  7. The Future of Research Symposium Boston 2015

    Type
    Blog Post
    ...is a STEM career counselor, Director of Education for the Jackson Laboratory for Genomic Medicine and a... successful and sustainable biomedical research sector at our first Future of Research symposium. The ...compulsory internship to be taken in an industry, regulatory, or clinical environment. It remains contested...the overall structure of the biomedical research sector, and what a tenable workforce structure may look...ideal and productive workforce look like for the sector – how many jobs are there for STEM graduates and...strong supporter of increasing the number of postdoctoral scientists at University of Maryland. Paula... at Scientific Meetings from Addgene Executive Director Joanne Kamens Learn how to be a successful manager...

  8. We’re Going Viral This Week!

    Type
    Blog Post
    ... be viral vector jokes and comics. Other new resources for researchers using viral vectors In addition...Addgene blog Download our Viral Vectors 101 eBook Find all of our viral vectors blog posts Learn more about...Starting today, our Twitter feed will be full of viral vector information. Each day, we’ll cover a different ...AddgeneGoesViral to find information on the basics of viral vectors, applications, and some new resources available...also investigates new strategies to improve viral vector production and data analysis. Our first paper came...

  9. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...in viral vectors, but easily fit in a TE. Transposons are also less likely than viral vectors to induce...DNA transposons are most commonly used in the laboratory for genome manipulation. When transposons are...transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a closer look at three different ... in human cells at rates similar to retroviral vectors. The hyperactive version of the SB transposase,...efficiency in human cells similar to retroviral vectors. The hyperactive PB transposase (hyPB) is ~10-fold...transfer Transposons are an alternative to viral vector delivery of transgenes, like for iPSC reprogramming... piggyBac, which is a big advantage over viral vectors (~5 kB payload for AAV and ~8 kB payload for lentivirus...

  10. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...Hendel et al. 2015). Low detection rates mean the investigator can be relatively certain that their samples...off-target mutations are essential data points for investigators looking to definitively link genotype and phenotype...Targeted amplicon sequencing doesn’t tell the whole story Figure 1: Schematic representations of the...viable options for multiplex sequencing in many laboratories. Nevertheless, unbiased detection isn’t as sensitive...genome-wide unbiased methods. However, most investigators use single cell clones for in vitro CRISPR experiments... Hough. We additionally thank Monica Sentmanat, Victor Dillard, and Ayokunmi Ajetunmobi for their contributions... Hough is a Biochemistry PhD candidate in the laboratory of Prof Steve Jackson at the Gurdon Institute...

  11. Design Tips for Prime Editing

    Type
    Blog Post
    ...nick, and monitor indel frequencies (Anzalone et al., 2019). Figure 6: Prime editors bound to ... Figure 4: Editing the PAM prevents the prime editor from re-engaging with DNA it has already edited...to pegRNAs drive editing (left), while prime editors bound to sgRNAs only drive nicking (right). Start ...opposite strand. Created with BioRender.com. MMR inhibitors PE4 and PE5: These approaches inhibit MMR to...inhibited. Created with BioRender.com. RNA protectors epegRNAs: epegRNAs include structured motifs...primer binding site (Nelson et al., 2022). PE7: This editor includes the fused RNA-binding protein La to protect...software, however. You may find it useful to ask a mentor what software tools they recommend — feel free ...

  12. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    ...cells survive their own secretions. Pro tip! Historically, colicin-producing plasmids have been referred...referred to as “Col plasmids” or “Col factors.” Virulence plasmids: microbial infection Virulence plasmids...other molecules to host organisms. For instance, vectors derived from the Agrobacterium Ti virulence plasmid... through direct contact. That’s conjugation! F factors can also be integrated into the bacterial chromosome...developing new cloning techniques based on these vectors (Shareck et al., 2004). Plasmid categorization...the many plasmid tools available in the Addgene repository. To learn more about some other common plasmid...Auriol, P., Vogel, T. M., & Simonet, P. (2001). Laboratory-scale evidence for lightning-mediated gene transfer...

  13. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...Washington Office Ben Corb - Director of Public Affairs at ASBMB Adam Fagen, PhD - Director, Early Career Scientist..., these issues which will continue to shape the sector throughout their careers (McDowell et al, F1000Research...working group (National Academy of Sciences. “The Postdoctoral Experience Revisited” 2014 Appendix B p93-95...funding and funding mechanisms require an engaged sector with effective advocacy skills. Advocating for ...Scientist Segment at AAAS Marnie Gelbart, PhD - Director of Programs at Personal Genetics Education Project...their respective fields: David Cameron, PhD - Director of Media Communications, Harvard University Ray...Advocate with the Perfect Science Talk Find a Good Mentor ...

  14. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...Diagnostics will play an important role in helping to monitor and slow this spread until vaccine programs can... switches for pathogen detection In 2014, as postdoctoral fellows with Jim Collins and Peng Yin at the.... The first was a new class of programmable riboregulators, called toehold switches, which allow for the...allowed gene circuits to be deployed out of the laboratory in a cell-free and biosafe format that was stable...circuit-based sensors to be used outside of the laboratory. The detection threshold for these RNA sequences...the technology through product development, the regulatory process, and manufacturing so that we can get...Leslie Dan Faculty of Pharmacy at the University of Toronto. His research is focused on developing cell-free...

  15. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...deliver in low-capacity vectors like AAV. AAV (adeno-associated viral vectors) are commonly used for in...Background Read About CRISPR Competitor NgAgo Learn How You Can Use piggyBac Vectors in Genome Editing Resources...preferential infection of certain tissues. However, these vectors have a low genetic capacity of ~4.5 kb. At 4.1 ...barely fits into AAV when you add in a gRNA and regulatory elements. To solve this problem, the Zhang lab...previously used SaCas9 (1053 aa/3.2 kb) with AAV vectors for efficient postnatal editing. LbCpf1 and AsCpf1...SpCas9. These nucleases should fit well into AAV vectors, especially given that Cpf1 gRNAs are also shorter...

  16. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    ..., 50ms “off”.  A pulse generator such as the A.M.P.I Master-9 Pulse Stimulator, which can control output...cable is threaded through—and fixed to—an optical insulator called a ferrule (below). The fiber optic cable...Channelrhodopsin 2 (Chr2) has been expressed in the motor cortex of a mouse. Upon connection of the ferrule...to give the animal the full range of motion, a commutator allows the patch cable to connect to the laser...University of Michigan and he is currently a Postdoctoral Fellow in the lab of Mary Jeanne Kreek at the... 1. Gutierrez DV, et al. Optogenetic control of motor coordination by Gi/o protein-coupled vertebrate ... neurons. Current protocols in neuroscience / editorial board, Jacqueline N Crawley  [et al]. 2012;Chapter...

  17. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...Keep the hood clean Hoods should not be used as storage cabinets. Clutter not only makes it difficult to...air conditioning on) so leave them in a sterile incubator. This also means that any medium or washing buffer...entering the hood and when putting them into the incubator). It also helps to get familiar with the morphology...results. Most cell culture media contain the pH indicator phenol red (pH range 6.0 (yellow) - 8.0 (red))...aliquot them into a number of vials, freeze them, and store all but one in liquid nitrogen. The frozen vials...master stock. This ensures that you have many cells stored in their original condition and can replace your...with the Agarose Method Plasmids 101: Mammalian Vectors Resources on Addgene.org Find mammalian plasmids...

  18. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    ... was contributed by Kusumika (Kushi) Mukherjee, editor of Trends in Pharmacological Sciences, a Cell Press...Shinya Yamanaka’s lab, where the reprogramming factors were delivered via retroviral infection [8]. The...Kushi) Mukherjee. Kusumika (Kushi) Mukherjee is the Editor of Trends in Pharmacological Sciences, a Cell Press...communication and publishing after completing her postdoctoral training from Massachusetts General Hospital...PMCID: PMC2752493. 4. Bongso, A. and M. Richards, History and perspective of stem cell research. Best Pract...embryonic and adult fibroblast cultures by defined factors. Cell, 2006. 126(4): p. 663-76. PubMed PMID: 16904174... cells from adult human fibroblasts by defined factors. Cell, 2007. 131(5): p. 861-72. PubMed PMID: 18035408...

  19. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ...development programs for bioscience educators. Using these hands-on tools, educators introduce their students to...This post was contributed by Tim Herman, director of the MSOE center for BioMolecular Modeling and the...dangerous. Subscribe to CRISPR Blog Posts How do educators and students use molecular models?   We created...something tangible. Under the guidance of a trained educator, these instances of focused attention become magical... their labs. Soon after we began working with educators in our Center for BioMolecular Modeling, we spun...our guest blogger Tim Herman! TIm Herman is the director of the MSOE center for BioMolecular Modeling and...Addgene.org Visit Our CRISPR Guide Page CRISPR/Cas9 History Browse All CRISPR Plasmids and Resources ...

  20. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...guest blogger Melanie Fox, founder and executive director of Central Indiana Science Outreach and a Postdoc...professional development I had a wonderfully supportive mentor and graduate office. I cannot stress how important...my blog helped immensely when CINSO started brainstorming the resources we wanted to provide online. ...in real time! I follow great online science communicators like Phil Plait @BadAstronomer, Emily Graslie...when I was trying to find local informal science educators, and ever since, the State Museum has been the...week. I gained experience explaining science to visitors of all ages, learned a lot about the dinosaurs...Melanie Fox is currently the founder and executive director of Central Indiana Science Outreach and a Postdoc...
Showing: 361 - 380 of 1082 results