...painstakingly cloned into the perfect empty backbone, made sure to add the right tags to your gene, and may have...provide enough background for you to distinguish the features of any common lab strain and determine whether... DNA, and more. Many mutations that make these features possible are present in most commercial strains...of protein expression in E. coli and the common features found in those bacteria. Check our our blog post...

...experimental results and 2) perform targeted off-target measurements at those sites to get an idea of how high the...effects, you still need to do your due diligence to ensure they are indeed a non-issue. Above we mentioned...these outcomes. Rigorous analysis is essential to ensure off-target events are below a given threshold for...your risk level is, how to lower it, and how to ensure off-targets don’t mess with your results when they...

... either category, but those are a subject for a future post! How are repressible promoters regulated...commonly used in yeast. In the first 24 hours of culture, when glucose is abundant and ethanol concentrations...during the late ethanol consumption phase of yeast culture. Repressible Binary Systems GAL4/UAS In Drosophila...the system, researchers have used split GAL4 architecture or combined the system with Flp/FRT, as reviewed...

...contest is over, we will host other contests in the future. Subscribe to the blog to stay in the know!   ... hunt works works: You will be trying to find pictures of Addgene pets with Addgene-branded clothing ...pet gives you a clue to help you find the next picture. If you find the next correct photo, you will see...

...Addgene gave to me: a method to capture promoter-enhancer 3D chromatin structures using CRISPR. On the eleventh... you’ve been enjoying our CRISPR blog series. Be sure to subscribe to the CRISPR blog posts so you don...

...flick the tube several times. This will help ensure ensure equal distribution. Lab tip #3: Barcode everything... the latest episode.  Have some suggestions for future tips?  Send your ideas to [email protected].  Additional...

... or hydroelectric in nature. The operation of -80C and -20C freezer temperatures, biosafety cabinet ventilation... lighting/HVAC requires a significant energy expenditure here at Addgene. These are critical systems that...

...key step to any experiment and can help prevent future headaches. To help you plan PEI transfections for...simple to make adjustments for the type of tissue culture dish and number of dishes used. Here’s to making... pHelper: 2 RepCap Select what type of tissue culture dish you will be performing the transfection in...

...What's a lab group in industry? The collaborative nature of working in industry and how delegating can empower...success. The importance of thinking about the big picture and not just the next experiment. Chapter 6: Advice... the treatment of various diseases, from heart failure to Alzheimer’s.  You can follow her on Twitter ...

...29:54 - 31:25 : Nature's defenses against gene drives. 31:26 - 33:08 : Plans for future work at the Wyss...advisory board where he will help shape Addgene's future. Listen to this episode for insights into the process...

...for years – searching our photo archives I found pictures back through 2009, though I suspect that our costume...months in advance. Our teams comb the news (pop culture events of the past year obviously make for good...takes home the ultimate prize – a glorious trophy. Sure it’s plastic and you have to share 1 trophy among...

...and proteins associated with pathogenesis. In his future work. Dr. Bengyella would like to use the expertise...to do excellent work in the biological sciences. Future awardees Now that this first round of awards has...enable scientists in developing regions in the near future. Additional Resources Apply to Recieve Equipment...

...what the founders hope to see in the repository’s future. If you’ve ever been curious about what it takes...success Chapter 6 - Developing Addgene’s Brand and culture Chapter 7 - Building Addgene as a global organization...Tufts Launchpad, and more Chapter 10 - Addgene's Future, the research team, and the importance of sharing...

...and look forward to hearing from her in the future. Future awardees We will be opening the Addgene/Seeding...them the most. Check back here to learn more about future awardees and their inspiring research. Additional...

...perform in vivo screens in other cell types. A key feature of this system is a lentiviral gRNA expression ...the statistical power of in vivo screens. Figure 1: Using FITS to perform in vivo CRISPR screens...clonal dynamics in lymph nodes (LN) and tumor cells. Figure used with permission from Milling and Markson et...

...also be expanded to include even more loci. Figure 1: Schematic overview of the all-in-one CRISPR/... work successfully to target multiple sites in cultured human cells. While it has not yet been demonstrated...dCas9-KRAB in our system,” Sakuma said. “These are sure to be powerful tools to modify the expression of...

...Today’s episode of the Addgene Podcast features guest host and Addgene European Outreach Scientist, Benoit...cooperation, and antibiotic production. Chapter 5: The future of Synthetic Biology. Wide adoption of synthetic...Biology and Innovation, Imperial College London and lectures synthetic biology for the Department of Bioengineering...

...University). Right there on the floor outside the lecture hall. It was wonderful to see the ad-hoc ‘sciencing...in vivo using ultrasound (for an example see the figure below). In addition to low costs, deep tissue penetration... imaging of microorganisms in mammalian hosts." Nature 553.7686 (2018): 86.PubMed PMID: 29300010. PubMed...

...more secrets - during a collaboration, members of Maureen Hanson’s and Rima Menassa's labs made the accidental...nm, photoconverted with 405 nm light, and then captured in the red state after excitation at 561 nm. This...conversion did not occur. Tobacco suspension cell culture with cytosol-targeted GFP. White circles represent...

...substantial amount of properly folded, relatively pure protein, but getting to this stage is often much... coli strains designed for protein expression. Cultures are then lysed and the protein of interest is ...disulfide bond formation. pCri also includes a special feature for difficult-to-purify membrane proteins. pCri...

...products is desired, as multiple selection markers can ensure that all desired tags have been integrated. Simply... to their genes of interest, allowing for easy capture and detection of proteins and complexes, without... single strain. In addition to the collections featured above, many other modular yeast tagging systems...

...progress towards understanding this disease. Figure 1: Scientists are continuing to do essential research...portal has recruited 3,218 Canadian volunteers, and features over 200 COVID-19 related research projects, funding... will remain an extremely important part of the future of research. To find other COVID-19 databases and...

...active in both E. coli and human cells. Using the structure of CasX at eight different states of assembly .... coli and human expression. Read the paper in Nature Find the CasX plasmids Drug inducible transcriptional...sequence-specific toxicity of Cas9. Read the paper in Nature Communications Find the Bikard E. coli Genome-wide...

...scientist in training can adopt to ensure opportunities in the future. Studies show that the majority of...an excellent resume ready to be submitted The structure and content of a resume can be crucial for a successful...

...proteins known to localize to specific cellular structures.The Davidson lab even included some of their own... see Supplemental Documents here). If you’re not sure where your protein of interest localizes in the ...easier for scientists everywhere to get a clearer picture of that intracellular world, even as the Davidson...

...This blog post is the first in a series that will feature advice for students, postdocs, and young Principal...their top 3 tips. Tom Ellis 1. Keep up with the literature. He recommends setting up a few Pubmed email ...“If you don't read, you're doomed.” 2. Always be sure to include various positive and negative controls...

...paper that appeared in Nature Biotechnology back in 2011 and then in another Nature Methods paper last year... far-red light. As the researchers described in Nature Communications last year, “The capability to control...

...proteins, DNA, and RNA used in CRISPR.   Figure 1: Overview of the parts of CRISPR. The bacterial...in the CRISPR array that will become guides for future targeted degradation. Before integration into the...CRISPR–Cas immunity and genome editing enzymes. Nature Reviews Microbiology, 20(11), 641–656. https://...

...archaea and eukaryotes such as yeast and plants. In nature, plasmids provide one or more functional benefits...for the life scientist or bioengineer.   Figure 1: Map of a plasmid with its elements described...constructed in the lab? Due to their artificial nature, lab plasmids are commonly referred to as “vectors...

...Zimbabwe Dr. Fiona Robertson (far left) is a Senior Lecturer and a Plant Molecular Biology and Biotechnology...manage insect pests and to support sustainable agriculture. For this project the expression of isolated .... Subscribe to the Addgene blog for updates and future collaborations. Many thanks to all of the awardees...

...of soundwave pressure required to collapse them: arg2 vesicles collapse at a lower pressure than arg1 vesicles...noninvasive imaging of microorganisms in mammalian hosts. Nature, 553, 86-90. PubMed PMID: 29300010. Additional ...

...get it into your plasmid backbone, and zap the mixture of DNA into cells. Unfortunately, not every cell...post, but don’t worry, we’ll cover screens in a future post. Positive selection: Cells that have gained...toxin genes, and the sacB gene). By tapping into nature’s toolbox, scientists are finding more and more ...

... distinct C-bend to escape possible danger (see figure). Larvae also perform spontaneous slow swimming...distinguish between swimming behaviors, the researchers measured the tail movements of 5 day old fish. The larvae...behavioural module in the vertebrate spinal cord." Nature 461.7262 (2009): 407. PubMed PMID: 19759620. PubMed...

...and a drug is developed, it’s good practice to measure the specificity of that target and validate the... is where CRISPR can save time and money to make sure quality candidates are selected.      CRISPR...discovery pipeline.   CRISPR and drugs in the future While CRISPR has already done a lot for drug discovery...

...in our repository and will discuss some of the features you may want to consider as you make your choice...vector that you will need. You will also have to make sure that your plasmid has been incorporated into the...promoter. Other vectors can be expressed in fly culture cells like S2 and carry the metallothionein promoter...

...advice from the successful PIs who were previously featured in "9 Tips to Achieve Success in Academia". Important...evaluated your plan? Have you read the relevant literature to understand where the field is today? Have ...the importance of keeping up-to-date with the literature of a couple of well-known journals that discuss...

...indefinitely at room temperature (thus also saving freezer space).  The procedure becomes less attractive... agar slices.  Molecular biologists used this procedure daily before the advent of techniques like PCR...

... is based on the Cas9 D10A nickase described in Figure 4 of the Cong, et. al, 2013 Science paper. For ...Restriction Fragment Length Polymorphism (RFLP) (see Figure 4 of the Cong, et. al, 2013 Science paper). If ...review articles, such as: Sander JD & Joung JK, Nature Biotech, 2 March 2014. Or browse the articles related...

...selective media, and provides those bacteria with a pressure to keep your plasmid. Viva la (bacterial) resistance... antibiotics against Gram negative bacteria. In future posts, we'll detail selection in non-bacterial ...strains have natural antibiotic resistances, so make sure your plasmid and E. coli strain are compatible! ...

...successfully reprogram. Unfortunately scRNA-seq only captures the gene expression of cells at a single time ...analysis and the remaining cells were maintained in culture. iEP cell physiology and reprogramming efficiency... lineage and identity in direct reprogramming." Nature 564.7735 (2018): 219. PubMed PMID: 30518857. Guo...

...the public in a meaningful way, and not solely to ensure a profit. Often, these companies believe they possess...chance to carry out a comprehensive review of the literature informing the notion of “heading” in soccer as... of my thesis: a survey instrument designed to measure how individuals respond to issues of unsettled ...

...the foundation of all good communication is making sure everyone’s on the same page. So for our first post...education is education that is provided through a structured environment such as the classroom or a defined...specific audience - giving consistency, purpose, structure, and intent to the messaging - while also creating...

...colleagues and is invested in improving research infrastructure, from our educational materials to our repository...and ScFvs, based on community demand. Figure 1: Antibody against PSD-95 from the Trimmer Lab...Reproducibility: Standardize antibodies used in research. Nature 518:27–29. https://doi.org/10.1038/518027a  Resources...

...seedlings or callus (undifferentiated plant cell culture), which is great for a wide variety of applications...found in all other plants, anthocyanins.   Figure 1: An adult plant expressing RUBY. Photos by Andrew...can be selected without antibiotics. (Top left) Mature silique (Arabidopsis fruit) with fertilized developing...

...stressors, including emotional stress or changes in temperature.  “The disease was first described in 1971,” ..., it is still sometimes wrongly diagnosed as a seizure disorder.” In 2012, researchers identified the ...Thailand will receive much better health care in the future based on our studies.” Thank you to all the BeHeard...

...that we aren’t still pitching for feedback.  Your venture should continuously evolve and respond to feedback...number of experts, including Harvard/MIT faculty, venture capital and angel investors, and business development...idea has potential to make a difference, then make sure you’re able to answer every possible question related...

...that are then barcoded for identification. This mixture of barcoded fragments is known as a *library*. ...additional benefit of plexWell™’s sequential tagging procedure, the quantities of library molecules for each ...multiplexed pool. The cost of these normalization steps, measured in time and consumables, can be daunting for large...

...addition to thorough ethical considerations, in making sure that the research has been adequately proven to ...unethical behavior (it’s hard to argue otherwise). To be sure, throughout the history of medical research, there...on necessary actions to review and decide on the future of genome editing technology. We must stop punting...

...formats. For example, several journals like eLife, Nature, and Cell use  Research Resource identifiers (RRIDs...resolvable compact identifier. Addgene works hard to ensure that materials available through our repository...Addgene Blog Read a blog post on Our Citations Feature Check out this post on Scientific Reproducibility...

... the library, use next-generation sequencing to ensure that the library has retained gRNA representation...produce high titer lentivirus from the library, ensure gRNA representation in the viral prep, infect target... you're interested in studying. You need to make sure your assay is scalable and robust. What you don'...