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We narrowed to 940 results for: Mpl;

Showing: 381 - 400 of 940 results

  1. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    ...specific examples. Instead of saying “I have excellent communication skills” try giving an example, like ...match the job requirements, by giving specific examples of your work and explaining how your experience...pointing to your transferable skills. In the above example, if you don’t have experience with ddPCR, but do...beginning is often the hardest part, here are some examples to get you started! Dear Dr./Professor [last ...strengthened an application. Even so, there were examples of great hires who hadn’t submitted a cover letter...

  2. Viral Production at Addgene

    Type
    Blog Post
    ...of the viral capsid proteins VP1, VP2, and VP3. Samples of viral preparations are subjected to polyacrylamide...proteins as a fraction of total protein present in the sample is also determined and used to determine purity...shown relative to the total protein present in the sample.   Sterility Viral vector preparations are ...rigorous barcode matching system that follows the sample from the DNA tube all the way to the cleared viral...throughout production, ensuring the identity of each sample from the DNA tube all the way to the viral preparation...Absolute determination of single-stranded and self-complementary adeno-associated viral vector genome titers ...

  3. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...mechanisms behind the cytotoxicity can vary. For example, the popular VSV-G envelope protein causes cells...are differentially inactivated by nonspecific complement mechanisms in serum. Sometimes this is due to...cells that the viral vectors were produced in. For example, production of vesicular stomatitis virus, HIV-...sugars end up in the envelope and are targets for complement-based killing by anti-alphaGal antibodies. The...good at infecting most cells, but not all. For example, LDLR expression in hematopoietic stem cells, T... as described above and is inactivated by the complement system in serum (DePolo et al., 2000). Other ...

  4. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...revolutionised the way we apprehend molecular complexes and is still a very useful tool.    Like a guitar...technique, but it is very important to follow some simple rules to avoid common pitfalls and to use it at... These issues can be overcome by choosing a complementary pair of fluorophores. To maximize the FRET signal...especially when you investigate unknown molecular complexes. But it is a parameter that you have to keep in...most efficient approach for your own system. The simplest and the most popular one is the sensitised emission...photobleaching method The acceptor photobleaching method is simple but limited to a single measurement. This method...

  5. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...locations using Cas9 and track the localization of the complex. Broccoli, the other green fluorescent aptamer,... RNA. Fluorescence is observed when these RNAs complex and the entire aptamer can hybridize and fold. ...uniqueness of the photostability of the Corn-DFHO complex. For dessert (last but not least), the only fruit-named...Other groups have used Mango to study RNA-protein complexes (Panchapakesan et al., 2017) and short non-coding...mango aptamer-fluorophore: a bright, high-affinity complex for RNA labeling and tracking." ACS chemical biology...transcription using a photostable RNA–fluorophore complex." Nature chemical biology 13.11 (2017): 1187. PubMed...

  6. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...endogenous homologous recombination enzymes can also accomplish the desired genomic modification. This technique... which results in a merodiploid. This can be accomplished by selecting for an antibiotic resistance marker...Valentovich 2014). We routinely clone the purified amplicons upstream and downstream of the desired deletion...included versions with gentamicin-resistance (amplified from TP997). We validated these vectors, which...scarless modification of bacterial genomes. It can accomplish large deletions, insertions, and even precise...system with lambda red recombineering enables simplified chromosomal gene replacement in Escherichia coli...

  7. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...multiple plasmids. For simplicity, we’ll use the second-generation system as an example.  The lentiviral ...double-stranded DNA transgene is incorporated into a large complex of both viral and cellular proteins and trafficked...the delivered payload is gradually reduced. For example, after using CRISPR/Cas9 to edit a gene, the expression...integrated piece to measure! Instead, two commonly employed methods for titering IDLVs include detecting vector...performing quantitative PCR titration of vector DNA in samples harvested 24 hours post-transduction (Wanisch and...

  8. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ...tough. Is the content accurate? Are we conveying complex ideas in the easiest way possible? Are the shots...Magic Pocket Cinema Camera 4K. It’s wonderful and simple to use, but when you need to juggle filming and...perfect height (Fig. 2). We recorded video using a simple, external video recorder and a mic clipped to her...video conferencing software. Interviewees could simply boot up Zoom and record themselves talking to their... and availability, a traditional solution was completely out of reach for Addgene.  So, onto research!...  Figure 4: The remote directing setup, complete with wireless video receiver, camera control, ...

  9. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...CRISPR may be the way to accomplish this tough job! Kamel Khalili’s lab at Temple University has demonstrated...the T-cell line 2D10. PCR amplification and Sanger sequencing of pooled samples showed that most of the ...undetectable in the blood. But the virus doesn’t completely disappear; it just hides in latently infected...the immune response. The second strategy is the simple removal of the HIV-1 genome from infected cells...

  10. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    ...postdocs. Unfortunately, even when universities have simple postdoc numbers sorted out there can be gaps in...Postdoc: An Institutional Perspective. This lack of complete information does not, however, prevent us from...scientifically minded, and able to communicate complex information - all traits you are likely to find...important take-away from this session was the idea that simply satisfying a percent requirement for diversity ...difficult question but not one that should be abandoned simply because it is difficult and is key to the continued...research. Panelist Moon Duchin provided a concrete example of how underrepresented individuals are discluded...

  11. What's New in CRISPR - May 2019

    Type
    Blog Post
    ...oligodeoxynucleotide HDR template covalently attached to the Cas9-gRNA RNP complex via the fused HUH endonuclease...genome-wide CRISPR inhibition Covalent tethering of DNA template to Cas9 SECURE base editors Nme2Cas9 CRISPR interference...inhibition library Covalent tethering of DNA repair template to Cas9 Homology-directed repair (HDR) can be ...Neisseria meningitidis, Nme2Cas9, that recognizes a simple dinucleotide PAM (N4CC), lending to high target...

  12. 6 Tips for Analyzing and Troubleshooting Sanger Sequencing Results

    Type
    Blog Post
    ...results Above (top right corner of this post) is an example of a seemingly clean DNA sequence (no Ns in sight...less and you might suspect contamination in your sample or consider asking your sequencing facility to ...to apply a special protocol for a difficult template. Anything more and you’re venturing into the uncertain...Use a silica spin column for purification of the samples you send for DNA sequencing If your sequencing ...requires you to perform your own Big Dye PCR amplification reaction (as opposed to using the all inclusive...

  13. Finding Your Perfect Job After University

    Type
    Blog Post
    ...This was particularly interesting as I had just completed my thesis on the study of e2Fb, which is a tumour...each advertised job received many applications, employers were requesting that only scientists with an MSc...cancer and I loved the practical aspects of testing samples in the lab, but I really struggled with the large...strong point. Working in industry: virology On completing my MSc, I again had to look at what type of work... solution attempted was more meetings, but this simply seemed to create more bureaucracy. As with any ...to a Work-Travel VISA, which means I can gain employment for up to 6 months in Australia or another country...

  14. Stepping into a Virtual Internship: My Experience at Addgene

    Type
    Blog Post
    ... to an internship on the journey to full-time employment. I have always been interested in the ways that...the forefront, no longer allowing myself to just simply exist without creating a storm. Given all the options...Canva, and I was able to create cute and on-brand templates for these posts. Some of recent Meet an ...what it was like to work at Addgene. This is exemplified in my work interviewing fellow Addgenies. I spoke...company that cares - not only in terms of its employees but in general about anyone who steps foot through...

  15. Management for Scientists: Seeking Feedback

    Type
    Blog Post
    ...from at least some employees. Watch out for "information allies" – these are employees in your group that...feel that giving feedback will be perceived as complaining. They may worry that it seems like "tattling"...have understood the speaker accurately. Thank employees who volunteer constructive feedback and suggestions...influenced by feedback. If you don't do this, employees will stop providing feedback pretty quickly. ...

  16. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...recombination that are typically employed during viral plasmid amplification (Fischer, KB. 2019). This means...expression profile at multiple steps from plasmid amplification in bacteria to transgene expression in vivo....of spontaneous reversion during AAV plasmid amplification. On the left homologous pairs of scrambled cre-recombinase... set of injections into your animal model. For example, in a recent study by the Wickersham lab (Lavin...essential that every new vector, whether it be a completely new transgene or merely a new lot, be thoroughly...

  17. CRISPR 101: Cas9 Nickase Design and Homology Directed Repair

    Type
    Blog Post
    ...this can be accomplished with either a PAM-in or PAM-out orientation. Just as the names imply, PAM-out designs...Overview of Cas9 nickase We’ll use SpCas9 nickases as examples for this post. The D10A mutation inactivates the...site rather than the optimal <10 nt distance. Employing two gRNA sites that are further away from the ...mutation site. Quick tips for nickase design Ready to employ nickases for your next CRISPR experiments? Here...

  18. SunTag and Fluorescent Imaging

    Type
    Blog Post
    ...this biological lesson and created a system to amplify fluorescent signals. Named for the "stellar explosion... However, this system is far from perfect. For example, If your fusion protein is expressed at low levels...expressed from a separate plasmid. This system amplifies the intensity of the fluorescent signal and enables...chance of phototoxicity Reduced photobleaching Simplified, long-term single-molecule tracking Caveats ...et al. “A protein-tagging system for signal amplification in gene expression and fluorescence imaging....

  19. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

    Type
    Blog Post
    ...medium containing the corresponding antibiotic. For example, if the DNA you’re inserting into the plasmid backbone...an essential amino acid), and a plasmid that complements the auxotrophy (i.e., synthesizes the critical... resistance spontaneously arises in yeast. For example, Markus Ralser’s lab created a yeast prototrophy...prototrophy kit containing 23 plasmids to complement auxotrophy. Negative selection: Cells that have lost a specific... gene. SacB can be used similarly to CcdB. For example, in the gene expression plasmid pGTvL1-SGC from...

  20. Antibodies 101: Fc Effector Functions

    Type
    Blog Post
    ...response to a viral infection a) antibody-dependent complement deposition (ADCD) b) antibody-dependent cellular... Erp, et al., 2019.    Antibody-dependent complement deposition (ADCD) Antibodies can bind to a pathogen... known as opsonization. In antibody-dependent complement deposition (ADCD), opsonization of IgG and IgM...pathogen activates the immune system's classical complement pathway, starting a cascade of events that results.... PMID: 10742152. Dunkelberger, J., Song, WC. Complement and its role in innate and adaptive immune responses...
Showing: 381 - 400 of 940 results