...about adeno-associated Viral Vectors (AAVs) before starting my graduate program at the University of Manitoba...and AAVs are non-enveloped viruses, so they are resistant to alcohol-based disinfectants. A 10% bleach solution...expression occurs quickly and results in long and stable expression (up to 9 months in mice [2]). Low Immune...tips that will help you think about how to get started with AAVs. Since my research is rooted in the CNS...Location, location, location! What do buying real estate and using AAVs have in common? The three most important... location, location! Just like this common real estate mantra, the choice of serotype (which determines... in the tissue/what cell type? In the CNS, for instance, there are marked differences among serotypes ...

...contributed by Laura Lee, a graduate student at Stanford University. Arabidopsis is a fantastic model organism...Agrobacterium tumeficans - a plant pathogenic species that stably integrates transfer DNA (tDNA) into the genomes... are also a variety of plant selection markers (basta, hygromycin, kananamycin, tunicamycin) (Nakagawa...plants. If your cultures have been incubated at stationary phase for too long, the cells in your culture...a colony from an LB plate. In my hands, a 5 mL starter culture inoculated from a colony on a plate will...density in 18-24 hours. I then add 0.5 mL of this starter culture to 100 mL fresh media, which is then ready...transformation in another 24 hours. The remaining starter culture can be used for colony PCR and glycerol...

...path.  A part of the career path story for most established grant professionals is learning to succeed in...often hear the term ‘grant writer’, but make no mistake, researching, winning, and effectively managing... with clear budget narratives, logic models, established partnerships and collaborations, and commitments...compliance with the grantor’s requirements as well as state and federal regulations. A grant professional’s ...no surprise that the U.S. Bureau of Labor and Statistics shows that employment of grant writers is expected... by 16% between 2010 and 2020. How did you get started in the grant profession? I asked Deborah Cook, ...on many different issues, and our program didn’t stay in one place within the city infrastructure. My ...

...guest blogger Sean Stacey! Sean D. Stacey is a Ph.D. student at East Tennessee State University. He works...This post was contributed by guest blogger Sean Stacey who recently attended both online and on site courses... online course had three aspects to it: videos starring Roberto Kolter, webinars taught by Peggy Cotter...were not too laborious, but helped build my understanding of how to generate a manuscript. At the end ...another email notifying me of my acceptance. I was ecstatic! I was going to become a serious scientific wordsmith...reinforced our newfound knowledge with practical understanding of how to apply it. The personal interaction...to get better. Well what does that mean? If you start a new technique, then type up the materials and ...

...after negative staining. Empty vector particles can be identified after negative staining and appear darker...generating a standard curve of known concentration. We regularly make and validate fresh plasmid standards. We...purposes in preclinical studies, it is important to understand what these values mean and how they can be compared...primer pair. Ct values vary based on the amount of starting material in the PCR because the sample must fall...fall within the linear region of the standard curve. In addition, high and low-concentration samples can..., which is a factor in AAV titration since the starting template is the purified viral vector solution...quantification by using the universal AAV Reference Standard Material (AAV SFM), which is an AAV sample that...

...discussing the review topic with your peers. Start reading! I started by reading other reviews because, as I...from a wide range of authors. Just start writing When I first started I thought I would read a bunch of... scientific review.” The second thing I did was stare in horror at the limited number of hits. So, here...outline the organization of the review Once you start reading, there will be a temptation to include every...normal day.” The way I broke this cycle was to just start writing. A word of advice: when reading, don’t feel... feel like you need to read every paper from start to finish. Find the information that’s relevant to ...results and contextualized them. No really, just start writing! Don’t worry about grammar or formatting...

...science majors, we do our best to understand the science. By understanding the hows and whys of our users...provides transparency to everyone involved about the status of each project. This creates accountability on...is our process for working on a new feature. We start with a story that describes the feature and what...collaborations between Addgene’s Product Team and the stakeholders (other Addgenies). Once we begin work on a story...closely with each other and with other teams. We start with writing demo details, step-by-step instructions...the Product Team who in turn collaborates with stakeholders to resolve the questions.  After the code is...from what I would have done. And finally the stakeholder accepts the new feature as done and we’re onto...

...on the cloning process, all constant regions, just the heavy chain constant regions or solely the Fc region...plasmid backbone containing all constant regions, just the heavy chain constant regions or solely the Fc region...great experimental plan that requires indirect staining of both targets in your specimen. You scour the...and heavy chains made up of variable regions and constant regions. Variable regions are unique to each antibody...antibody and convey specificity to the target while constant regions are identical for antibodies within certain...rat) are cloned into a backbone expressing the constant regions from an alternative isotype (purple, mouse...for multiplexing. In the case above, you can’t costain with your desired antibodies because the pair belong...

...in Table 2 is a standard and should always be included—the live/dead stain. This stain is in fact not ...in your analysis. Lastly, there are fixable dye stains that react with free amines on and inside the cells...antibody-fluorophores, fluorescent reporters, and/or dye stains used in your flow cytometry experiment are known...promote electrons within atoms to a higher energy state. This process is called excitation. After a short... period, the electrons revert to a lower energy state, whereby a photon is emitted. This step is called...laser but only "bright" for the blue laser. To understand why that is, let's look at Figure 2. The blue...markers with any fluorophore. Bleeding Before we start building your first panel, we'll talk about one ...

...guest blogger Sean Stacey! Sean D. Stacey is a Ph.D. student at East Tennessee State University. He works...This post was contributed by guest blogger Sean D. Stacey. Opinions on the apps discussed in the article ...great for lab life. These apps prevent you from constantly searching the interwebs for lab resources, and...computer. Lab Archives will help both you and your lab stay organized allowing you to upload anything from protocols... you. 2. Gibco Cell Culture by Gibco Whenever I start to learn a new technique, I try to look for a resource...for a mass spectrometry app. I digress. When I started learning proper cell culture technique, I tried...manual. Overall, it’s a great tool for someone starting cell culture work. 3. NEB Tools by New England...

...undergrad working with cancer cell lines. As I started planning out my own experiments in graduate school... As a busy researcher, you’ve probably already started coming up with time management techniques of your...load yourself up with too many tasks. If you’re constantly switching between different projects, you might...make notes on as I progress. I’ve also recently started using Trello to manage larger, multi-person projects.... Knowing this, I hand write to-do lists at the start of each day to remind myself of the specific activities...where you’re pulled in a thousand directions at the start of the day, writing your to-do list the night before... each project and how useful each item is. For instance, if I don’t have draft blog post ready for publication...

...Nature report from researchers at Amgen, it was stated that only 11% of the academic research in the literature...due, at least in part, to insufficient and non-standard record keeping despite scientist’s best efforts...The familiar paper-based lab notebook is still a standard approach to record-keeping in many labs. In a ...functionalities that are hard to imagine on paper, such as instant search through all saved data and automated writing...writing of reports. To be able to understand the situation in depth and provide the solution that fits ...adoption within academic environments to better understand the issues labs are facing in terms of budget...to ELN adoption. This includes financial outlay, staff hours, troubleshooting, and the fact that long-term...

...them survive under severely cold conditions. To understand their unique importance, let’s begin with a few...while before we can firmly gain a handle on understanding and appreciating their biology. The importance...of processes. The food processing industry, for instance, employs psychrophilic enzymes for milk processing...proteins for industrial or research purposes requires stable host cells (the simplest example would be a transformed...studied, and they are convenient to maintain under standard conditions. However, one of the many proteins ...conditions of ~0°C, which drastically affects their stable folding and efficient functioning. But what about...haloplanktis Trigger Factor), that would allow the stable folding and expression of recombinant psychrophilic...

...Eligibility check Scrutinize every opportunity prior to starting an application to assess your eligibility, interest...share a common set of criteria such as citizenship status, academic year, and grades that serve as automated...can be found on the Addgene blog.   3. Personal statement brilliance In addition to sending in a cover letter...letter, you are always asked to draft a personal statement, which is the perfect opportunity to paint a comprehensive...devoting some time to writing a solid personal statement will pay dividends. This is the only opportunity...tools that can help you craft a solid personal statement.  4. Letters of recommendation wisdom  Reach out...for the character you painted in your personal statement. Make certain to choose your recommenders based...

...from the Institut Gustave Roussy. Professor Kroemer discussed the effects of starvation on cancer therapy...tumor-associated inflammation for therapeutic benefit ”as stated by the organizers. My overall impression from... that it is too fragmented and focused ... to understand cancer and defeat it we need to attack it from...treatment (Ref 1). As described in the presentation, starvation or the use of caloric restriction mimetics trigger...hepatocellular carcinoma, a cancer which can develop from a state of chronic hepatitis caused by hypernutrition and...consumption. Chronic inflammation is a dangerous state for the body to be in and usually self-limiting ...effects kick in trying to resolve the inflammatory state by inducing immunosuppressive measures, for example...

... the internet, but start by downloading my handy list of suggestions.  Get started. Have regular meetings... other as you transition to the next stage in your careers! Stay in touch…this group will be the core ...elusive teachers, supporters and advisors? I tried to start a more formal mentoring program at my company, but...mentoring relationships.   If you would like to start a group mentoring program at your organization, ...provide a step-by-step, how-to guide to get one started. Advantages of the group mentoring format Adding... speaking to grad students and postdocs I have started to talk to trainees more about peer mentoring. ...Because the participants are at a similar development stage, they have a lot to learn from each other. Some ...

...Communication blog series, you’ll hear from Susan Keown, a staff writer at the non-profit Fred Hutchinson Cancer ...Center. Susan Keown’s science communication journey started with something unexpected: termites. “I was really...says Susan. Her passion for insects lead her to start a PhD in entomology at the University of Maryland...’m like, “Oh my god, what can I do?! This is a mistake! Why did I even come [to grad school]!” He asked...know if she liked science writing. She decided to start a blog to test it out and discovered she not only...continued working for Palladian Partners remotely, but started looking for a new job because “it kinda sucked ...hired a full-time social media manager. “I usually start out my days looking through our Twitter feeds and...

...that is resistant to the toxic effects of CcdB. Another commercially available, CcdB-resistant strain ...components that are key to the cloning reaction. Understanding the specific components is essential for choosing...actually contain your insert of interest. Simply stated, ccdB makes cloning easier by selecting against...But exactly how does ccdB accomplish this? Let’s start with a brief history of the gene and how molecular...non-recombinant vectors (re-ligated empty vectors, for instance) would still express ccdB and therefore would ... investigator’s gene of interest.   CcdB resistant E. coli strains complete the system But ccdB alone...contains the F plasmid (F’ strains) will also be resistant to CcdB, as the native ccdA will be present. Most...

...definitely help in understanding if your strategy will be successful, and to avoid easy mistakes that could affect... and the expected result is due to a PCR mistake or a mistake by the DNA analysis software. Now that you've...hopefully obtain your coveted plasmid with no substantial delays. Design, design, design Before diving ...many softwares out there than can help you at this stage and that can be used to simulate in silico cloning... of insert compared to the backbone plasmid; a starting point could be a molar ratio of 1:2 (plasmid:insert...antibiotic in the plate correspond to the antibiotic resistance marker present in your plasmid. You should also... mix in different bacterial strains (stbl2, NEB stable, etc.), and/or use a different temperature (30°...

... at MIT) where he worked on understanding bacterial antibiotic resistance. During this time, Dr. Koeris...In the third installment in our podcast series, we chat with new Addgene Board Member, Michael Koeris...this idea, Drs. Koeris and Lu founded the biotech startup company, Novophage. As you'll hear Dr. Koeris explain... decided to develop phage for use in a biotech startup 12:16 - 15:15 : Why Sample 6 began to develop tests...Koeris to personally take on the risk of founding a startup company? 32:20 - 34:37 : Whose responsibility is...Sample 6 website Check out Dr. Koeris' Website, biotechstart.org Read Sample 6's Paper on Bacteriophage-based...Other Posts in Our Podcast Series Get Additional Startup Advice Learn How to Be A Good Manager from Addgene...