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Showing: 661 - 680 of 1019 results
  1. Tips for Titering Your Lentiviral Preps

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    Blog Post
    ...options as well as the benefits and drawbacks of different methods (for comprehensive protocols for all of...used in the transfection; if preparing several different lentiviruses in parallel, this approach should...that standard conditions be used when comparing different batches of virus. In addition, some viral preps...step of a successful transduction experiment. References 1. Kutner, Robert H., Xian-Yang Zhang, and Jakob...
  2. Plasmids 101: Degron Tags

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    Blog Post
    ...you can make an informed choice! References and Resources References Bondeson, D.P., Mullin-Bernstein,...are titratable. With these tags, protein levels aren’t limited to ‘on’ and ‘off’; varied levels of drug...profile their effectiveness with over a dozen different proteins. In the end, you can’t predict with absolute...
  3. 10 Basic tips for mammalian cell culture

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    Blog Post
    ...Also, keep all liquid containers closed when they aren’t in immediate use. This will prevent spills, but...could change their growth rates and morphology, rendering their continued use inadvisable for experiments...in a lab where multiple people are working on different things, this will help you avoid cross-contamination... Immunology. You can find Sana on LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling...
  4. xCas9: Engineering a CRISPR Variant with PAM Flexibility

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    Blog Post
    ...species, like S. aureus and N. meningitidis require different PAM sites than SpCas9, in this case, NNGRRT and...single PAM recognition. Created with BioRender.com. After 24 days of phage propagation, Hu ...percentages, with fold changes compared to SpCas9 in parentheses. The development of xCas9 3.6 and 3.7 shows ...is working for you in the comments section! References Hu, Johnny H., et al. "Evolved Cas9 variants ...
  5. Plasmids 101: Optimizing Plasmid Yields

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    Blog Post
    ... Let it incubate Check the bacterial strain Differences between E. coli host strains can impact plasmid...might want to include an extra wash step or use a different strain. For most applications, Addgene uses and...inadequate plasmid DNA yields. Why would volume make a difference? In the simplest of terms, more volume = more...conditions play a critical role in plasmid DNA yield. Different plasmids and strains will vary in their optimal...
  6. Anatomy of a Plasmid Page at Addgene

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    Blog Post
    ... ever found yourself bamboozled by all of the different kinds of information on our plasmid pages? Well...depositing lab, although you may have an entirely different use in mind! This section of our plasmid pages... you to use in your Materials and Methods and References sections when citing the use of your colleague...e.g. if it was derived from a plasmid made in a different lab - or one obtained from Addgene!).  It also...
  7. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

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    Blog Post
    ...anymore. The kit contains sequences for several different fluorescent proteins. Visualizing fluorescence...use our promoters and terminators, but then they aren’t sure how to convert their own coding sequence to...with the Athena SWAN Charter and it has really strengthened the network of female scientists. Twitter has...for those who go to work in cancer biology.  References: A Golden Gate Modular Cloning Toolbox for Plants...
  8. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...alleles, or introduce a premature stop codon to render an RNA nonfunctional. In their quest to build a...are present in the gRNA spacer, as ADAR will preferentially edit an adenine if the template has a cytosine...(Abudayyeh et al., 2019). To avoid some of the inherent drawbacks for RNA editing of natural cytidine ... Valley contributed to updating this article. References Cox, David B.T., et al. “RNA editing with CRISPR-Cas13...
  9. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ...known quantities of DNA. Additionally, an AAV reference, a virus with a known titer, can be used to confirm...both results would be valid. This is why the AAV reference is critical. It allows you to determine if your... PCR (ddPCR) does not require a standard or a reference, meaning saving on reagents (and time)! AAV titering...becoming a popular option for PCR experiments.  References Furuta-Hanawa, Birei, Teruhide Yamaguchi, and...
  10. Viral Vectors 101: The Retroviral Lifecycle

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    Blog Post
    ...dimerized linear ssRNA strands (which can have a different genomic sequence from one another) — by various... with lentiviral replication.  Resources and References More resources on the Addgene blog Viral Vectors...popular lentiviral plasmids γ-Retrovirus Guide References Cullen, B. R. (1992). Mechanism of action of ...Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives. Viruses, 13(7), 1288. https...
  11. Isolating Genomic Regions of Interest with the CRISPR System

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    Blog Post
    ... adaptations of this system have been made in different fields of study. In addition to the identification...purification with an anti-Cas9 antibody. These different tag systems and affinity purification schemes ...dystrophy (FSHD), a disease for which there is currently no cure (Campbell et al., 2018). Find plasmids...please also see our interview with Dr. Fujii. References Campbell, Amy E., et al. "NuRD and CAF-1-mediated...
  12. Lentiviral Vector Uses and Overview

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    Blog Post
    ... applications. Read on to learn more about the current (and future) applications of lentiviral vectors...facilitates nuclear export of transcripts). A different viral envelope protein (Env), usually from VSV-G...plasmid number. For more information on the key differences between the 2nd and 3rd generation packaging ...use lentivirus in the comments section below! References 1. In vivo gene delivery and stable transduction...
  13. Tips for Screening with Yeast Two Hybrid Systems

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    Blog Post
    ...when separated (2), and that DBD and TAs from different systems could be combined effectively (3). In ...a more vigourous screen is to use a number of different bait and prey vectors. This has been shown to ...split ubiquitin system has been devised (8). References 1. Casadaban M.J., Martinez-Arias A., Shapira...Science. 1986; 231:699-704. PubMed PMID: 3080805. 3. Brent, R., and Ptashne, M. A eukaryotic transcriptional...
  14. Cpf1: A New Tool for CRISPR Genome Editing

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    Blog Post
    ...endonuclease domain, indicating that Cpf1 functions differently. Since Cpf1 loci are widely distributed across...could be adapted for genome editing. The use of a different nuclease could potentially overcome some of Cas9...Figure from Kleinstiver et al., 2016. Cpf1’s strength may lie in its low off-target editing rates, determined...shown the diversity of CRISPR-based systems. References 1. Zetsche, Bernd, et al. "Cpf1 is a single RNA-guided...
  15. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    .... While these differences may seem subtle they result in a key physiological difference between lenti-... system is that it involves transfecting four different plasmids into the producer cells (two packaging...as the viral long terminal repeats (LTRs) are different. As a result, lentiviral and retroviral packaging...
  16. Negotiating Work and Life: How to Find the Joy

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    Blog Post
    ... to a conference. Really. No one will starve while you are away a few days. Also, your children will enjoy... yourself, “I’m not married,” or “I don’t have children, so my life will be under control.” Think again... hard not to do this. For a family with working parent(s), the sooner your kids can make a decent meal...
  17. Hot Plasmids: Fall 2024

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    Blog Post
    ...middle of proteins) and performs well in many different antibody-based applications.  IPI’s recombinant...virus 5 and their use in revealing antigenic differences between human, canine and simian isolates. J ...systems don’t always work across species due to differences in natural DNA repair pathways and spontaneous...at focal adhesions in living cells in several different contexts.   Figure 5: Left: Mechanism ...
  18. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ...tracking experiments (Rizzo, 2009). With many different FPs that emit across the visible spectrum, researchers...of existing FP and epitope tags, researchers in Loren Looger’s lab have developed new molecular tags combining...antibodies. The “spaghetti monster” name is a reference to the deity of the social movement, the church... to the expanded use of smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance...
  19. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

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    Blog Post
    ... Optimizing FLP FRT through smart engineering Different FLP versions – making FLP efficient The first ...efficiency to Cre (Kranz et al. 2010). These differences in recombinase efficiency are important to keep...should be visualized at the same time from two different plasmids this approach becomes problematic as ...the genOway press release for more information. References 1. Atasoy, D., Y. Aponte, H. H. Su, and S. M....
Showing: 661 - 680 of 1019 results