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...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...

...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...

...opportunities where I could make mistakes and learn from them. I started at my previous company by taking...transitioning from academia to industry is whether to stay hands-on in the lab or pivot to a managerial role...mystery at first! Image used under CC.    Understanding the transition While a PhD equips you with exceptional...burdensome task of explaining the steps to a lab assistant. However, I recognized the need to shift my mindset...needed to adapt based on individual needs. For instance, I had one colleague who preferred detailed instructions...instructions and regular check-ins, which helped them stay on track and feel supported. Another team member...in the basics of management. Invest time in understanding key principles through books, workshops, and...

...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...

...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...

... 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...

...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°...

...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...

... 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...

...present. Using a Standard Curve To do that calculation, you’ll need to include a standard curve in your ...your standard curve to plot to a linear equation (remember y = mx+b?). Figure 5: A standard curve...that relies heavily on antibodies is ELISA, which stands for Enzyme-linked Immunosorbent Assay. ELISA are...readers can detect is 4.0, so if you max out your standard curve and it reaches an asymptote, you won’t be...wells in that range of optical densities. If your standard curve does not have a linear portion or the OD...when you repeat the experiment by diluting your standard curve, performing a serial dilution of your experimental...Once you’ve identified the linear portion of your standard curve, you can calculate the concentration of ...

...designs and has improved protein yields and stability. This stabilized spike protein can be used for the development...LH,  Begeman A, Stark JC, Shon DJ, Fox DM, Puschnik AS, Bertozzi CR, Carette JE, Stanley SA, Harris E, ...Apr 30, 2020 and last updated Jul 12, 2021. What started out as a small collection of plasmids for coronavirus... of cathepsin S, L, and B. This became a great starting point for SARS-CoV-2 researchers to investigate...Calabrese (activation), from Addgene to better understand which human genes are important for SARS-CoV-... packaging) and contain an IRES-Puro marker for stable cell clone selection. For more information on these...JS (2020) Structure-based design of prefusion-stabilized SARS-CoV-2 spikes. Science 369:1501–1505. https...

...9514755. Richard Gronostajski: NFI-related diseases of neural development Richard Gronostajski is a professor...Paula Videira, Michael McMurray, and Richard Gronostajski who each are being awarded free plasmids form...disorders of glycosylation Paula Videira is an Assistant Professor at the Faculty of Sciences of NOVA University...Applied Molecular Biosciences (UCIBIO) focused on understanding the physiological and pathological roles of ...biosynthetic pathway and have a wide range of clinical manifestations. PMM2-CDG is the most frequent CDG type characterized...University of Buffalo. His research focuses on understanding the structural determinants of protein-DNA interactions...they affect brain and muscle development. The Gronostajski lab has generated mice wherein it's possible...

...existed! Addgene recently started automating a few protocols to make standard lab processes easier and ...compared to a lab assisstant filling about 200 vials in 45 minutes by hand. How can you start automating your...robot and to find out how you can get one. As a starting point, I would recommend conferences like Lab ...majority of Addgene plasmids are shipped as agar stabs in glass vials. Have you ever wondered who fills...fills all those vials? Well, it used to be a lab assistant, but with increased plasmid demand it was too ...can do to get into the robotics spirit include: Start using electronic multichannel and single channel...Addgene would love to move forward and automate. For instance, vial labeling would be much easier with a Tube...

...a high standard of sequence quality across all plasmids in our collection. This standardization provides...Even the most high-fidelity polymerases can make mistakes and lead to the accumulation of errors in DNA ...as they were deposited before 2017 when Addgene started sequencing all incoming plasmids. If you are interested...Addgene’s QC team. Email [email protected] to get started! Addgene’s QC process Addgene prioritizes the sequence... to ensure all plasmids pass our high-quality standards and contain the expected plasmid features.  ...QC team updates and reviews our sequencing QC standards to ensure the entire team is using the same rubric...replacement for the plasmid, and the full QC process is started over again after receiving the replacement plasmid...

...deconstruct and understand this complexity using existing C. elegans mutants. For instance, we use a mutant...research on worm locomotion, but our worms are constantly moving so we need good ways to monitor and track... C. elegans molecular biology are based on the standard worm genome. While C. elegans are often thought...and so far most of their funding has come from kickstarter and fundrazr projects). This allows the project... to a wider group of people. This is a perfect starter project for our computer scientists, many of whom...interested in biology but don’t really know how to start applying their skills to it. The Kittybiome project...Funding Makerspaces One of the main challenges when starting a new project at a makerspace is finding the financial...

...number is then determined by comparing results to a standard curve. For CNV assays, it is recommended to use... (Ma, 2014.) As with all qPCR assays, samples, standard curves, and normal controls must be run in triplicate... the variance seen within multiple runs of the standard curve. Signaling molecules: Sybr Green and Taqman...noise, due to the large amount of dsDNA in most starting materials, and is not particularly sensitive. ...each reaction, math happens (this time, Poisson statistics), and the readout gives an absolute value of ...concentration values, rather than by comparison to a standard curve, and most results can be confidently interpreted...Bell, 2018.) Even though it does not require a standard curve or a normal control, it can take a few hours...

...plants, and agricultural crops in particular, resistant to disease. They also devise biosensors designed...that they work well in plants and give similar stability to transcripts as the ones that people use all...all the basic and necessary elements in order to start using the MoClo system. The paper also gives quantitative...have any particular tips for those looking to get started with MoClo? Patron: The parts will be familiar ...the parts that you need from the kit and getting started making your own constructs with Golden Gate MoClo...conceptual barrier. Sometimes people don’t know how to start. They might want to use our promoters and terminators...they don’t have any prejudice about it. They just start and go ahead. I can see science speeding up. Addgene...

...verify the starting methionine of a gene but the blastx results start the alignment at a more distal amino ... available, we primarily use Standard Nucleotide BLAST (blastn), Standard Protein BLAST (blastp), and ...Translated BLAST (blastx). NCBI has a terrific getting-started guide for BLAST, which includes a simple explanation...differences below. Optimizing blastn searches On the Standard Nucleotide BLAST page, the first decision to make...homologous regions, which by default are ignored in a standard blastx search. An alignment omitting repeated ...the sequencing result, we often choose between a Standard Protein BLAST (blastp) and blastx search to verify...sequence alignments that BLAST is unable to handle, Clustal is our frequent choice for pairwise or multiple...

..., such as the starting material (DNA-based vector or purified RNA) and the initial state of the dsRNA ...experiment functions as the building block of our understanding of complex biological processes. Many tools ... knockouts have long been considered the “gold-standard” for LOF studies, the high gene copy number found...function due to the use of exon skipping, alternative start codons, and/or the phenomenon of transcriptional...transcriptional adaptation (Smits et al., 2019; Sztal and Stainier, 2020). Additionally, a study that directly compared...approaches and considerations present at the first stage of an RNAi-based investigation. Creation of the ...gene silencing efficacy, thereby requiring a substantial time investment. Once the target mRNA is cleaved...