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We narrowed to 1,027 results for: LEA

Showing: 221 - 240 of 1027 results

  1. Transferable Skills Guide: Public Speaking

    Type
    Blog Post
    ... in front of an audience and convey your ideas clearly and strategically is powerful in almost any career...public speaking, and I'm happy to share what I've learned along the way. Own your time When I was an undergrad... your purpose The purpose of our assignment was clear: present a paper to our classmates so they could...don’t just happen to you or me (though, let's be clear, they have all happened to me.) They happen to everyone...speaking as a scientist is that you will have to learn this skill in its most difficult form. You’re presenting...audience or not in your own native language. And, at least in the biomedical field, with very little text on... speaking as a scientist is that you will have learned this skill in its most difficult form, which makes...

  2. CRISPR Cheat Sheet

    Type
    Blog Post
    ... CRISPR nuclease. Note: Most CRISPR plasmids from Addgene produce either a gRNA, a nuclease, or both...Editors: CRISPR nucleases that cut or modify RNA as opposed to DNA. Fusing the nuclease to an adenosine...CRISPR system to cut other DNA or RNA sequences. Nucleases: Proteins that bind to gRNAs and are directed ...sequence. Base Editors: Modified versions of CRISPR nucleases that make single base changes in the DNA sequence...Activate/Repress: Modified versions of CRISPR nucleases that can’t cut DNA (dCas9, for example) that are...gene. Screen: A library of gRNAs with a CRISPR nuclease target multiple genes in a cell population. The...

  3. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...Resources on the Addgene Blog Learn More about Homology Directed Repair Learn More about Non-Homologous End...editing team at the Allen Institute for Cell Science. Learn more by visiting the Allen Cell Explorer at allencell.org...useful coding sequence (i.e. GFP) into a gene while leaving other features of the gene intact. Here, we show...based on known functional aspects of the protein gleaned from the literature and engagement with researchers... Institute for Cell Science Plasmid Collection, please visit the Cell Catalog on the Allen Cell Explorer...hiPS cell lines generated using these plasmids, please visit the Cell Catalog on the Allen Cell Explorer... study cellular structures and processes and can lead to the better understanding of cell biology. We ...

  4. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    ...Finish Your Thesis” online course.  To learn more about her site please visit www.FinishYourThesis.com.   ...all of the above categories. In addition, I also learned about two other kinds of difficult supervisors:... their minds. When you come to a meeting with a clear agenda, you will immediately get your supervisor...supervisor to support you. 3) Explain the problem and leave your emotions out of the discussion Do not assume...a high likelihood of miscommunication, which can lead to even more conflicts. If your supervisor is a ...clam), you will probably need support from your colleagues or other supervisors to complete your work. Whether...package your challenges with your supervisor into learning opportunities as you are advancing career, you...

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

    Type
    Blog Post
    ...MOIs. The presence of the 428 bp Surveyor nuclease cleavage product indicates a gene edit. GFP gRNA-transduced...basis of the Surveyor assay is that Surveyor nuclease cleaves double stranded DNA (dsDNA) at basepair mismatches...mismatches at the site of the edit. Using Surveyor nuclease followed by gel electrophoresis, these mismatches...mismatches are indicated by the presence of DNA cleavage products. The appearance of the 428 bp band indicates...I designed (listed above) worked well and I got clear bands using the conditions above. 2. Design your...product, you would still be able to identify the cleavage product. 3. The Surveyor kit had a lot of good...Before digesting the DNA duplexes with Surveyor nuclease, save some of the duplex and run it as an undigested...

  6. Synthetic Biology, Artificial Chromosomes, iGEM & More - Interview with Tom Ellis

    Type
    Blog Post
    ...world's leading yeast researchers in the growing field of synthetic biology and is the UK lead of the ...Benoit to interview Addgene Advisory Board Member and lead researcher at Imperial College London, Dr. Tom Ellis...construct a synthetic yeast chromosome. Listen to learn more about Dr. Ellis, synthetic biology, synthetic...advice for anyone pursuing a career in academia. Learning how to build a lab and a research program and ... thanks to our guest, Tom Ellis! Dr. Ellis is a lead researcher at the Centre for Synthetic Biology and...    Additional Resources from the Addgene Blog Learn about the Synthetic Biology Education Nonprofit ...Nonprofit BioBuilder Plasmids 101: Golden Gate Cloning Learn About Working at a SynBio Startup in Our Interview...

  7. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ... so if you are interested in learning more about any one of them, please click on the links throughout...with a special emphasis on these five categories. Please note that many of the following cloning methodologies...ends. These fragments are then combined with an exonuclease, a polymerase, and a DNA ligase to carry out ...FastCloning, first described by Chaokun Li and colleagues. Here you simply PCR-amplify your vector and ...When mixed, topoisomerase I provides both the endonuclease and ligase function while 5’ adenosine overhangs...additional enzymes required. FastCloning may be the least expensive of all the options discussed as it only..., as one tube of enzyme should be enough for at least 50 cloning reactions. The ligase cost is comparable...

  8. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...University of Zurich), especially with my deep learning-wizard colleague Ahmed Allam. Continued training led to...desired edit, it simply becomes impossible (or at least unreasonable) to test them all. This is where PRIDICT...efficiency and building prediction tools with machine learning (PRIDICT, PRIDICT2.0).   How to apply PRIDICT...prediction tool (ePRIDICT). In summary, machine learning-based prime editing prediction models (such as...biology. His expertise includes applying machine learning techniques to make sense of biology, with a special...SpCas9 activity prediction by DeepSpCas9, a deep learning–based model with high generalization performance... editing efficiency and product purity by deep learning. Nature Biotechnology, 41, 1151–1159. https://...

  9. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...the IRES element, scientists have adapted "self-cleaving" 2A peptides into their muliticistronic vectors...mulitple genes from the same mRNA. The term "self-cleaving" is not entirely accurate, as these peptides are...peptide bond at the C-terminus of a 2A element, leading to separation between the end of the 2A sequence...sequence and the next peptide downstream.4 The "cleavage" occurs between the Glycine and Proline residues found... common 2A peptides employed by scientists. 2A cleavage is universal in eukaryotic cells, and, although...although some scientists report close to 100% cleavage with some of these peptides, no consensus has been reached... added to the 5' end of the peptide to improve cleavage efficiency. How do I get started? If you are looking...

  10. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    ...bioinformatics and HTS, we all had much to learn. And, many wanted to learn more, even those not directly involved...continue such discussions. Beyond the excitement of learning about new cutting edge science, researchers of...in journal clubs. They also have likely had the pleasure of attending both productive and not so productive... (HTS) data we generate and maintain. It became clear that while several people across teams had various...same characteristics as a successful meeting: a clear purpose and effective time management, for starters...feedback you receive include the sense that people learned something new? Did the meeting begin and end on... published science. They also serve as a way to learn about work outside of one's own field in a structured...

  11. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    ...based on the experiments of Kim and colleagues using zinc finger nucleases to harness non-homologous end joining... (DSB) approach at a variety of loci. We were pleasantly surprised by the efficiency of the method. One...polymorphism analysis, allele-specific PCR, T7EN1 cleavage assay, sequencing, RT-qPCR, or immunoblotting ...genetic configurations. Watch the following video to learn Daniel and Matthew's procedures. The video details...genetics of blood cell development and disease. Learn more at his webpage, bauerlab.org.     Matthew Canver...chromosomal deletions in human cells using zinc finger nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. ... interspaced palindromic repeats (CRISPR)/Cas9 nuclease system in mammalian cells. J.Biol.Chem. 2014 Aug...

  12. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...with other proteins or nucleic acids. Read on to learn more about this versatile technique.  Immunoprecipitation... only to the Fc portion of the immunoglobulin, leaving the antigen binding sites free to capture the target...conformations and interactions. For difficult to release proteins, including those in the nucleus, more ... an IP, as proteins in an IP are not denatured (learn more about antibodies here!) When using Protein ... immobilize the antibody, it is recommended to preclear the sample to remove any proteins that may interact...nonspecifically with the capture antibody or Protein A/G. To preclear, incubate the sample as you would in an IP using...interactions from residual non-specific proteins but leaves the target protein/capture antibody intact. The...

  13. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

    Type
    Blog Post
    ...to the site and activates its nuclease function. Once bound, TnpB cleaves the DNA at a position 15–21 base...sequence of commonly used nucleases like Cas9. This is where TnpB offers a clear advantage. At approximately.... Among the most well-known CRISPR-associated nucleases are Cas9 and Cas12a, the heavyweights in the genome... has become a key focus in recent years. TnpB nucleases represent an exciting class of compact genome ...proteins. Functionally, while Cas9 contains two nuclease domains (HNH and RuvC-like), and Cas12a has one...strand), while TnpB, like Cas12a, uses just one to cleave both strands. Similar to how Cas proteins require...increasing the overall size), using TnpB as the core nuclease could help keep the total construct size smaller...

  14. Antibodies 101: Affinity Tags

    Type
    Blog Post
    ...applications. Tag cleavage must be performed after primary purification and requires a second clean up step to...protease cleavage at sites immediately following the tag. Unlike MBP, the GST tag can be cleaved off from...need for an extra clean up step of the tag – a handy feature. If you decide not to cleave the GST tag after...Waugh, 2002). The tag can also be proteolytically cleaved after purification, so as not to perturb protein...to remove the cleaved tag from solution. MBP antibodies are available for detection and/or purification...

  15. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ...ideas in the easiest way possible? Are the shots clear enough for the audience? As Addgene’s Science Video...SynBio, it was time to start our Design-Build-Test-Learn cycles. If you’re not familiar with this, it’s a...process!   Figure 1: The Design-Build-Test-Learn cycle adapted for video making.   Cycle One:... pulled it off! Check out the PCR Primer video! Learn: Producing videos like this would prove to be difficult...phones. Test: I loved the end result, but it was clear this wasn’t going to work for our other videos. ...1280x720 resolution at best. Some came in even lower.  Learn: While this works well for social media videos, ...chat with me remotely via an audio chat (Fig. 4).  Learn: This works great for on-camera presentations and...

  16. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...CRISPR Featured Topic Page Learn about Non-Homologous End Joining Learn about Base Editing Read 3...can be caused by intracellular factors such as nucleases and reactive oxygen species, or external forces...: The 5’ DNA end of the break is resected by nucleases to create a ssDNA 3’ overhang. This will serve... a primer for DNA repair synthesis, ultimately leading to the formation of double Holliday junctions (...second DSB end. The individual HJs are resolved via cleavage in one of two ways - each junction resolution ...single HJ forms during this process and resolves by cleavage of the crossed strand. While this pathway may ...point to note is that Cas enzymes can continue to cleave DNA if the gRNA/PAM site remains intact. In other...

  17. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...different promoters followed by leader sequences (encoding small leader peptides) downstream of which you... been used to date . If you have any questions, please write to us in the comments section.  FX cloning...gene of interest. Translation of the standardized leader peptide prevents secondary structures in the mRNA...Shine-Delgarno sequence found at the 3’ end of the leader sequence. Using a variety of GFP and RFP fusions...Katarzyna, et al. "Small changes in enzyme function can lead to surprisingly large fitness effects during adaptive...reported in the literature include RNA detection, RNA cleavage, and translational repression or activation: ...Choudhury, Rajarshi, et al. "Engineering RNA endonucleases with customized sequence specificities." Nature...

  18. Intro to Our Summer SciComm Series!

    Type
    Blog Post
    ...classroom or a defined learning program. Learners cannot (theoretically, at least) fully and freely engage... fall under informal learning; however, many others partner with formal learning to engage students at...education that is provided, typically ad hoc, for learners who can choose to engage or disengage with a very..., and their expectations of you, consistent and clear.   And now that we’re all on the same page, we’re...re ready to get into the more exciting parts - learning more about science communication itself! See you...

  19. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    ...applications. We recently released a new variant of jGCaMP7 (keep reading) and will be releasing more variants in...viral vectors. Here are some of the AAV we have released so far in 2019. You can also browse our entire...pAAV-mDlx-NLS-mRuby2 CAG-NLS-GFP pAAV-hSyn-mCherry Read on to learn more!  Calcium sensor AAV The GFP-based calcium...inhibitors that respond to blue light. We recently released a constitutive (pAAV-Syn-Archon1-KGC-GFP-ER2) ...coming soon! These vectors listed below should be released at Addgene in the next 6 months, pending quality... particular catalog item is available for order please email [email protected] with the vector name and...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

  20. Fluorescent Biosensors for Measuring Autophagic Flux

    Type
    Blog Post
    ...This decrease in pH quenches SEP but not DsRed, leading to emission of only red fluorescence. The ratio...ratio of red to green fluorescence allows for clear distinction between vacuolar/lysosomal and cytoplasmic...lysosomes. Therefore, the formation of autophagosomes leads to an increase of yellow puncta (GFF positive/RFP...autophagosome with a lysosome. An increase in autophagy leads to more yellow and red puncta, while a block at ...at fusion of autophagosomes with lysosomes leads to an increase in yellow puncta but a decrease in red ...Addgene! GFP-LC3-RFP-LC3∆G: A fluorescent probe that releases an internal control Another autophagy biosensor... following translation, this fusion protein is cleaved to generate one molecule of GFP-tagged LC3 for ...
Showing: 221 - 240 of 1027 results