...format that was stable for up to a year at room temperature (Pardee et al, 2014). As a demonstration of the...the project by initially designing and ordering several dozen versions of the toehold switches and amplification...Addgene As we report, we have embedded this next generation of toehold switch-based RNA sensors within an...

...in AT-rich genomes. Additionally, whereas Cas9 generates blunt ends 3 nt upstream of the PAM site, Cpf1...with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches at gRNA positions 1, 8, 9, and 19-23...sticky end” transgene insertion mediated by Cpf1-generated overhangs. Exploring the efficiency of these two...

...and mutant loxP sites has allowed scientists to leverage this system in new, creative ways. Today’s post...potential to express the transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic...Double-floxed inverse Orientation), FLEx systems are generally considered "Cre-On" meaning that your gene of ...

... the right choice. I spoke with our Financial Operations Specialist, Lucy Hammer, about why she did not...question came from Lucy Hammer, our Financial Operations Specialist. She said, “I’ve found the most purpose...the office. Main takeaways from Meet an Addgenie Overall, the Meet an Addgenie project was one of the biggest...

... and biological. Even the most high-fidelity polymerases can make mistakes and lead to the accumulation...populations. All of these topics require additional considerations to resolve. Sequencing and assembly issues ...acquired from the MiSeq for assembly or with enough coverage over plasmid regions to confirm their presence...

...select the components they want for each module, generate the vector file, and quickly clone their vector... Step 4: Naming your assembled plasmid and generating the vector file on the Fragmid website. ...4, is to give the assembled plasmid a name and generate the vector file. Ta-da! Most (180/181) Fragmid...

...drawn to the idea of working as a team to develop therapies that could directly impact patients. And, to be...beginning of March, and the jobs are mixed into the general careers section of their web site. Each position... and is applying this towards researching neurodegenerative diseases.   Additional resources on the Addgene...

...specific recombination outcome using PCR and then generate a monoclonal cell line with the floxed exon. You...B, A Limaye, & AB Kulkarni. (2009) "Overview: Generation of Gene Knockout Mice". Curr Protoc Cell Biol...Basics of Homology Directed Repair Read about Generating Knockouts with CRISPR through Non-Homologous ...

... modulators, and chimeric effectors. Nucleases generate a classic double-stranded break, while modulators...with researchers all over the world. That’s an average of 27,272 plasmids distributed per year, or approximately...it clear that it’s also a story of community, cooperation, and sharing. Sharing this story is a truly special...

... Prevotella and Francisella), was annotated in several genomes and is classified as a type V CRISPR system...Again, they were in for a surprise! Whereas Cas9 generates blunt ends 3 nt upstream of the PAM site, Cpf1...with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches at gRNA positions 1, 8, 9, and 19-23...

...quick tips to get things going: If your PTA has a general meeting and a means to request funding for events...and think of a way to make this section more interactive. If you can think of a good analogy that involves...but I also tried not to be an alarmist.  The overarching guideline for all of these questions, and for...

...connection levels and interaction. Can bring in things to be physically interacted with. Public speaking...s much more challenging to design a hands-on interactive activity using elephants than it is to design...

...needed for EM is not something you find in the average cell and molecular biology lab, and EM is limited... is able to expand evenly in all directions to several times its original size. Depending on the protocol...depend upon the specific gel recipe used, ambient temperatures, incubation times, and the physical properties...

...deprioritize genes that could be good targets for therapeutic development, particularly in cancers that are... ancestry which will reduce the likelihood of generating genetic dependency data with an ancestry-based...researching ancestry groups. And, of course, it’s imperative for scientists to be aware of and actively use...

...the grantor’s requirements as well as state and federal regulations. A grant professional’s job duties ...Dr. Cook served as a principal investigator for several research grants. One of the benefits of being a...management, and program evaluation are highly transferable. Use your experience and your passion(s) to ...

...sites differ at the nucleotide level but share an overall structure of 13 base pair palindromic repeats separated...efficient The first version of FLP discovered has a temperature optimum of 30 °C and is therefore inefficient...led to the identification of FLPe which has a temperature optimum of 37 °C (Buchholz, Angrand, and Stewart...

...are below the new exemption level (the current average postdoc salary is estimated at around $45,000, ...tests must be met. In the Department of Labor’s general guide to the effect on higher education, you can...Perez, Fair Pay for Postdocs: Why We Support New Federal Overtime Rules, made clear that institutional arguments...

...scientific mentor is typically a professor who operates a research group. Within that research group, ...everything! The bare minimum should be taking notes on general protocol steps and other tips and tricks your mentor...my dissections go smoother next time?” or more generally “How am I doing? Can you tell me what’s going ...

... with low affinity for ligands. When integrins interact with extracellular ligands, their ectodomains ...sites and do not interfere with integrin function. Overall, the synthetic rAbs demonstrate remarkable specificity...cells. What’s next IPI is working with Addgene to accelerate biomedical research by distributing antibodies...

...still go horribly awry when several factors aren’t taken into consideration. 1. Always focus on the science...equipment that you use in your current lab first. Generally, major equipment can be defined as equipment with...

...with a 2:1 BSc in Molecular Biology (roughly a B average in the American system), in 2009 (at the beginning...structured and controlled. I worked from Standard Operating Procedures (SOP’s) and every item I tested was...sustainable. After working at this company for several years, I was ready for a change.  Working abroad...

...resonance energy transfer (FRET) is a biophysical interaction between two fluorophores. When the first fluorophore...& Zhang, 2006), quantifying antibody/antigen interactions (Saraheimo et al., 2013), and visualizing structural...different biomolecules you’d like to test for interaction. In fact, targeting a FRET fluorophore to a biomolecule...

...multiplex gRNAs, some being: Using dual nickases to generate a knockout or edit. This can help reduce off-target...gRNAs, with four being the ideal number. First you generate four unique kanamycin-resistant plasmids, each...each unused promoter. This step is necessary to generate all of the overhangs needed for the final ligation...

...Toolbox for quickly generating zebrafish transgenic lines to study neurodegenerative diseases. Listen to...

...restriction digested with PstI or EcoRV, these plasmids generate 100 bp or 1 kb DNA ladders, respectively. Unlike.... L., & Tan, S. (2017). The pPSU Plasmids for Generating DNA Molecular Weight Markers. Scientific Reports...

...outlets meaning you’ll have the opportunity to interact with and discuss your work with many different...applying CRISPR for mouse genome editing. Post: Generating Mouse Models Using CRISPR/Cas9   Erik L. Snapp...

...hydroelectric in nature. The operation of -80C and -20C freezer temperatures, biosafety cabinet ventilation...

...viral lots. Aligning with Addgene’s mission to “accelerate research and discovery by improving access to...calculations on how to prepare a plasmid stock used to generate a standard curve, visit the AAV Titration by qPCR...

...department at MIT. Crystal now works on protein therapeutics at Novartis - listen to learn more about Crystal...in industry. The importance of recognizing and leveraging your unique skills set while also adapting to...

...Genetics Institute in Cambridge, MA and Sequana Therapeutics in San Diego, CA. She earned a B.S in Biology...Addgene as the Chief Financial Officer and Chief Operating Officer. Prior to Addgene, Kenneth was an investment...