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  1. 9 tips for a successful postdoctoral experience

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    Blog Post
    ...searched for and eventually identified a postdoctoral mentor. A good mentor typically writes the grants that ... of reference- the graduate school mentor, the postdoctoral mentor, and at least one more person that ...was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at theJanelia Research ...reason most people enter postdoctoral training. Similarly, there are several factors to consider when selecting... of your postdoctoral training experience. If you're going to commit to doing a postdoctoral fellowship... you how to write as well as mentor. Yet, even the most dedicated mentor is not always available for mentees...different roles? A second or third mentor. Therefore, seek out additional mentors. I found this beneficial for...
  2. Adenoviral Vector Production and Troubleshooting

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    Blog Post
    ...Adenoviral vectors (AdV) are attractive vectors for research applications and gene therapy: they can ...Three generations of adenoviral vectors: the first generation vectors are deficient in the E1 and E3 regions... second generation vectors are deficient in E1, E2, E3, and E4. Gutless Ad vectors (helper-dependent (...creating adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the transgene...Adenovirus type 5. Three generations of adenoviral vectors The adenovirus virion is a non-enveloped particle...medicine (2017).   1st Gen AdVs are stripped of regulatory genes E1 and E3. Without these genes, AdVs cannot...lysate is your primary low titer rAdV-S, and can be stored at -80C for later use or used immediately for amplification...
  3. Viral Vectors 101: Pseudotyping

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    Blog Post
    ...lentiviral vectors, you need an envelope protein on the virus’s surface and a corresponding receptor in the...lentiviral vector will infect, pseudotyping can help. In pseudotyping, you produce viral vectors using viral...glycoprotein. Lentiviral vectors are also great at infecting neurons and cause minimal inflammatory response, but...Sandoval S (2017) Pseudotyped Lentiviral Vectors: One Vector, Many Guises. Human Gene Therapy Methods ...the host cell. Some of these envelope-receptor pairings are broad, allowing delivery into many cell types...virus. How to pseudotype To pseudotype your viral vector, you use the same protocol for generating lentivirus...also be cell type dependent: if a cell has few receptors for a specific envelope protein, it takes up less...
  4. Viral Vectors 101: Transductions

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    Blog Post
    ...Production at Addgene Viral Vectors 101: What is a Virus? Viral Vectors 101: Viral Vector Elements ...nucleic acid cargo of your choice. First, a viral vector is selected to perform the transduction with: either...desired DNA to be integrated can be cloned into the vector that you have selected. The nucleic acid will be...produced, and viral particles are packaged, when the vector is expressed. Once the design and construction ... it’s time to make virus! The constructed viral vector containing the cargo of interest is then typically...layers of risk management when working with viral vectors. Typically, one day after transfection into the...variety of broadly applicable and popular viral vectors. Check out our viral selections to see if we have...
  5. Beginner's Guide to Viral Vectors

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    Blog Post
    ...use viral vectors for many experimental purposes. To help you make sense of all the viral vector information...Subscribe to Viral Vector Blog Posts from Addgene Need more information on viral vectors? Check out our ...our viral vector guide pages, our Lentivirus FAQ, and our viral service pages. Click on the Image Above ... viral vector...
  6. Editor's Choice, October 2016

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    Blog Post
    ... Read All of Our Editor's Choice Blog Posts Last month there was plenty of excitement at Addgene. Our...Increased Postdoc Wages,” Future of Research executive director, Gary McDowell, gives a thorough yet concise rundown...Grad School Advice Part 2: Building Community  My editor’s choice pick for this month is “Grad School Advice... editor's choice...
  7. Editor's Choice, September 2016

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    Blog Post
    ... Read All of Our Editor's Choice Blog Posts As I’m sitting in the San Francisco International Airport...surpassed 60,000 views for the month of September! Historically we do better in September than in the summer...for FOP, Running for AJ I have two picks for the editor’s choice posts this month. The first, "Technique...contributed by Kurt Swanson, husband of Addgene Director of Biology, Lianna Swanson, and researcher at ... editor's choice...
  8. Editor's Choice, August 2016

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    Blog Post
    ...In response to a bit of a Twitter storm surrounding NgAgo, its creator, Han Chunyu of Hebei University ... Read All Our Editor's Choice Blog Posts The Addgene Blog has had quite the up-tick in views over the...by PCR & Single Base Editing with CRISPR For my editor’s choice picks this month, I’m picking two posts...give a great science talk from Addgene Executive Director Joanne Kamens. I highly recommend taking a look...Transductions Single Base Editing with CRISPR Cas9 Activators: A Practical Guide 10 Steps to a Perfect Science...
  9. Editor's Choice, July 2016

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    Blog Post
    ...each and every month, we've decided to start an "Editor's Choice" series. Each month, I'll summarize the...on topics ranging from new tools you can use to monitor cellular perturbations to how CRISPR has made it...I love all of our blog posts, my pick for the "editor's choice" post this month is from guest blogger ... editor's choice...
  10. Lentiviral Vector Uses and Overview

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    Blog Post
    ...future) applications of lentiviral vectors! Lentiviral vector history and components In the early 1990s...design a viral vector system that could do both, Addgene depositor Didier Trono and collaborators turned to...Lentiviral vectors are one of the most popular and useful viral vectors in the lab. Advantages of lentivirus...dividing and non-dividing cells. Lentiviral vectors are the vector of choice for many CRISPR applications,...viral vector systems based on retroviruses like Moloney murine leukemia virus (MMLV). The vectors could...adeno-associated viral vectors can also target non-dividing cells, lentiviral vectors can carry much more...Lentiviral Plasmid FAQs. Lentiviral vectors in the lab Lentiviral vectors are incredibly popular in the lab...
  11. Plasmids 101: FLEx Vectors

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    Blog Post
    ...Genome Engineering Using FLEx AAV Vectors? Learn about Viral Vector Elements Read Our Primer on Optogenetics...improvement of optogenetic adeno-associated virus (AAV) vectors. Listed below are a few additional uses for FLEx...and Ghyselinck NB. A directional strategy for monitoring Cre-mediated recombination at the cellular level...PMC3406316. Find FLEx plasmids from these Addgene depositors: Ed Boyden Bernardo Sabatini Wenbiao Chen Liquin...
  12. Plasmids 101: Multicistronic Vectors

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    Blog Post
    ...due to factors such as the experimental cell type and the specific genes cloned into the vector.3  2A ... the creation of bicistronic or multicistronic vectors. Unlike promoters which will create unique mRNA...for each gene that is expressed, multicistronic vectors simultaneously express two or more separate proteins...blog post we’ll cover basics of multicistronic vectors: why they are useful, how they work, and how to...get started with them. Why Use multicistronic vectors? Detecting cells that are expressing your gene,...expressing your gene of interest at high levels. Unlike vectors that express screenable or selectable markers from... the same transcript. Of course multicistronic vectors do not have to exclusively be used as a means of...
  13. Plasmids 101: Mammalian Vectors

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    Blog Post
    ...using synthetic vectors and cultured mammalian cells. Of course, these mammalian vectors must be compatible...Instead, reporter genes are often employed to easily monitor transfection efficiencies and expression levels... mammalian vectors...
  14. Plasmids 101: Yeast Vectors

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    Blog Post
    ...elements for vector propagation. This post, along with a future companion post on mammalian vectors, will catch...typically found as a single copy. Unlike the ARS vectors, CEN vectors are stable without integration. Yeast Episomal...plasmid maintenance within an E. coli cell, but vectors can be widely utilized across many different cell...core replication and resistance features of yeast vectors and explain how they differ from the bacterial ...previously described. Why do scientists use yeast vectors? Yeast are eukaryotes and thus contain complex ... and yeast cells. To be useful in the lab, the vectors must contain a yeast-specific origin of replication...replication of fission yeast, Schizosaccharomyces pombe, vectors at the end.) In a previous post, we briefly discussed...
  15. Choosing a Good Mentor for Scientists

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    Blog Post
    ...Science Mentors series: Part 1: "What Makes a Good Mentor?" and 6 More FAQs About Science Mentoring ...choose an advisor/principal investigator (PI) that will also serve as a good mentor. To read more about what...choosing a lab for postdoctoral work. However, selecting a lab with a PI who is a good mentor is important ...to learn about her mentoring style. How can I find out if an advisor is a good mentor? You have to ask ...on this topic), for this second entry in the “Mentoring for Scientists Series”, let’s focus on how to ...what makes a good mentor, see the previous post in this series. Check out Joanne's Reddit AMA Different...Different types of mentors for grad school vs. postdoc Choosing an advisor to complete a successful PhD is different...
  16. Deciphering the Mysteries of Behavior with Viral Vectors

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    Blog Post
    ...confirms the previously suggested inhibitory mechanism of locomotor sensitization and might open new ways...studies using viral vectors in behavioral neuroscience. With the ability of viral vectors to fine tune gene...all of our viral vectors blog posts Find tips for getting started with viral vectors for behavioral neuroscience...is activating, inhibiting, or modulating. Viral vectors, in contrast, are considerably more versatile and...behavioral tests, but now we’ll focus on how viral vectors have changed the scope and power of three commonly...tests To study and quantify locomotion, scientists monitor an animal’s movement in an open field box or a ...particles expressing an activating chemogentic receptor (hM3Dq) into a subset of neurons within the LC...
  17. Antibodies 101: Buffers, Storage, and Conjugates

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    Blog Post
    ...outside factors that need to be considered as well. In this post, we’ll touch on antibody storage, buffer....  Storage   Once you get your exciting new antibody, the second thing you’ll want to do is store it ...what temperature to store your antibodies at, but it also matters what they’re stored in - their buffer....the way, you can put your antibody in short term storage (think days to weeks) in the 4°C. If you’ll only...primarily used in Western blots and ELISAs, cannot be stored in sodium azide at all, as it inhibits HRP. In ...excitation by a laser, which is then read by a detector panel in the machine. Fluorophores are light sensitive...sensitive, so antibodies conjugated to them need to be stored either in a dark, opaque vial or wrapped in aluminum...
  18. Viral Vectors 101: The Retroviral Lifecycle

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    Blog Post
    ...blog Viral Vectors 101: Viral Vector Elements Viral Vectors 101: Types of Viruses Viral Vectors 101: Viral...would not be great for a gene therapy vector! For viral vectors, only the protein for reverse transcriptase...components of the vector particle, some of the RT protein gets packaged inside of the vector particle, along...and this is where viral vectors and natural viruses diverge. For the viral vector, its job as a tool is ...Central PMCID: PMC372876 Dong, W., & Kantor, B. (2021). Lentiviral Vectors for Delivery of Gene-Editing Systems...seem like an unlikely candidate to use as a viral vector, but retroviruses have been developed into some...gamma-retroviruses — that have been developed into retroviral vectors and 2.) a subclass of retroviruses called the lentiviruses...
  19. Viral Vectors 101: Optogenetic Tools

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    Blog Post
    ...Bicistronic vectors combine excitation and inhibition by using both excitatory and inhibitory channels activated...BioRender.com.   Opto-G-protein coupled receptors  G-protein coupled receptors (GPCR) regulate cellular function...some broad classes of optogenetic tools, and what factors to consider when choosing one.     Figure...light-gated ion channels, optogenetic G-protein coupled receptors (OptoXRs), and photoswitches. Light-gated ion ...stimulation to either activate or inhibit a specific receptor or pathway.  Light-gated ion channels Light activated...light within a specific range of wavelengths. Excitatory channels allow positively charged ions such as...gradient to depolarize it (Nagel et al., 2003). Inhibitory opsins are usually light-driven pumps that pass...
  20. Viral Vectors 101: Virus Safety

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    Blog Post
    ... on the Addgene blog Viral Vectors 101: Types of Viruses Viral Vector Elements Tips for Preventing... contact with than GFP. Similarly, if the viral vector carries an shRNA to a gene essential for cellular...cellular function, this would also be a higher risk vector. While it is unlikely that you, or anyone else ... you can receive appropriate treatment and/or monitoring. Virus-specific concerns   In nature, some viruses..., some adenovirus strains are known to cause respiratory diseases in humans, earning this virus a BSL-...
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