...coupling of Cre-lox with FLP-FRT recombination to generate additional recombination events, and the synthesis...and sea anemone mKate2.) They then successfully generated custom antibodies to each of these proteins and...analysis. To achieve even cell labeling, they generated farnesylated derivatives of the fluorescent proteins...previously visible with Brainbow-1 and -2. The general structure of Brainbow-1.0 is retained in Brainbow...constructs are stably maintained through at least six generations. In addition to the neuronal pThy1-Brainbow constructs...

...twice the length of the monomer (~11.6 kb). Image generated by SnapGene software.  Thanks to increasing...which has been used in plasmid vaccine and gene therapy development to examine plasmid topologies (e.g....stationary phase. Growing the culture at a lower temperature such as 30°C or optimizing the incubation time...Fishel RA, James AA, Kolodner R. recA-independent general genetic recombination of plasmids. Nature. 1981...294184a0. PMID: 7029308. Goebel W, Helinski DR. Generation of higher multiple circular DNA forms in bacteria...

... would thrive on this level of attention and interaction (and I did). Some of the information you will...advisor. They are always traveling, or too busy to interact regularly with the newer lab members. You must...careers?    Will you be encouraged to develop transferable skills by attending career events or being involved...your own choosing? Does the PI keep up on the literature and pass on relevant papers? Will you have access...resources on the Addgene blog Check out our transferable skills guide Learn about careers in science ...

...started by designing a novel topology, then iteratively generated 3D models and amino acid sequences that ...topology of protein” diagrams, which show the interactions between amino acids in an approximate, two-dimensional...crosslinked peptides proposed as a scaffold for a new generation of drugs, all developed with their protein design...

...ways of generating antibodies, for example through the use of less toxic adjuvants; 2) generating complex... The minimal fees from distribution funds our operation. Antibodies are glycoproteins that are part of...increased avidity to high profile antigens; and 4) generating mAbs against new cancer biomarkers. We believe...

...) (Miller et al., 2005). Although this tight interaction is not covalent, later work showed that introducing...ER and mitochondria reveals dynamic organelle interactions. a) Labeling scheme. b) COS-7 cell ER labeled...without any cloning at all! As mentioned above, several variants of these tags have been developed over...https://doi.org/10.1021/cb900062k Gautier, A., Juillerat, A., Heinis, C., Corrêa, I. R., Kindermann, M.... Pick, H., Vogel, H., & Johnsson, K. (2003). A general method for the covalent labeling of fusion proteins...

...not right for the development of phage-based therapeutics. After a brief stint at the venture capital ...Novophage's focus. The company went from developing therapeutics to developing phage-based tools for detecting...develop tests for food products as opposed to therapeutics 15:15 - 16:26 : Means for testing for pathogens...for Dr. Koeris to transition from developing therapeutics to developing a diagnostic? 36:53 - 38:00 : ...

...are required to ensure that you get full library coverage in pooled library screens. Availability and Use...metalloproteinase-9 secretion, and identifying lysine methyltransferase substrates. We’re incredibly happy that lentiCRISPR...of the many plasmids being distributed to and accelerating research in labs all around the world. We thank...Browse Our CRISPR Posts Learn about Using CRISPR to Generate Knockouts Read Some Common Ways to Deliver CRISPR...

...of different conformations and thus potentially interact with and alter the proteins they label. In fact...chaperone Ydj1, both of which display sophisticated interactions with multiple other proteins. Again, little ...indicating that they compromised protein-protein interactions. If you’re looking to a tag a protein, inntags...tags of choice for examining protein-protein interactions, particularly in single-molecule or high-throughput...

... the key component of most CRISPR systems and generates double strange breaks at target sites 5' of the...your 20 nucleotide guide sequence of choice (for generation of a gRNA that will target SpCas9 to the approrpriate... expression post transfection by western blot. Overall, this is a versatile mammalian expression vector...Browse Our CRISPR Posts Learn about Using CRISPR to Generate Knockouts Read Some Common Ways to Deliver CRISPR...

...DayCon. DayCon is a one-day conference aimed at the general public that provides an accessible introduction...conference-style experience that was accessible to the general public both in terms of cost and content level....of disease, cancer, and viral infections. An interactive lunch with student presenters and board members... of shorter, more narrowly focused lectures, interactive panels, and a student poster session. All DayCon...

...available at Addgene, also gives the option to stack several expression cassettes onto a single T-DNA in subsequent...rounds, dramatically reducing the time required to generate multi-construct transgenic plants. In the most...choice, BsaI, which can be very sensitive to temperature fluctuations. He says their lab learned the hard...into the public domain in the process. “We had several motivations for making GreenGate available through...

...towards the objective is collected and focused onto a camera for visualization. Illuminating a sample in the...is excited at one time and captured in a single camera exposure. This is much faster than relying on raster...fast, sensitive detector such as a scientific CMOS camera. If plasma membrane dynamics are the main focus... excitation occurs through the fluorophore’s interaction with an evanescent energy field. Lightsheet microscopes... to a higher photon flux via the more efficient camera detector and lack of a light-reducing pinhole. ...

...s how science progresses after all, with each generation of scientists standing on the shoulders of their...Technology Policy issued a memorandum calling on Federal agencies with over $100 million in annual R&D expenditures...support increased public access to the results of federally-funded research. 2)      In June, the White House...Bohannon published an article describing a sting operation in which he submitted a phony paper to a list ...

...then formulate a plan to further develop your transferable skills and reach your career goals.      Although...subject matter discussed at the meeting, but the overall tone of the meeting is almost more important. As...Wunderlich Z, Estrada J, Park J, Bragdon MD, Lopez Rivera F, Biette KM, DePace AH. Mol Cell. 2015 Jun 4;58... What Makes a Good Manager Anyway Developing Transferable Skills During Science Training ...

... the interactive photo graffiti wall. I had a lot of fun playing the prototype of an interactive game ...about her grandfather’s humble beginnings as an operator of a printing press. Just as her grandfather used...the newspaper, Addgene uses E. coli bacteria to generate copies of plasmids. The community of scientists...

...opportunities, protocols, publications, and more to accelerate the pace of research into COVID-19. While this...opportunities, and community response initiatives. Accelerate the translational impact of research Universities...important for creating research synergies and accelerating the translational impact of research activities...Alliance at the CCRP2. Other groups have focused on iterating on methods for coronavirus research, sharing resources...

...from the standard UAS/GAL4 system is toxic to proliferating cells. A Cre-estrogen receptor ligand binding...about new recombinases suitable for this system. Gerald Rubin’s lab sought to make complex genome modifications...specific recombination site for each enzyme, were generated to test recombination efficiency. When expressed...PubMed. Reduction of Cre recombinase toxicity in proliferating Drosophila cells by estrogen-dependent activity...

...and Dr. Paul Stoffels from J&J. The panel was moderated by a non-scientist from Bloomberg news. Given ...who shared first authorship on their seminal, cooperative paper in Science.  Interestingly, this project...finding they have been able to use CRISPR/Cas9 to generate mice with genes knocked out in the insulin signaling..., open academic exploration in this area will accelerate the launch of clinical applications. During ...

... brand new Europe office located in London and operated in collaboration with LGC Standards. And now you... VII. During my years in research, I developed several new plasmids, including: plasmids to study the ...and shared by others to develop an anti-tumour therapy using antibodies targeting dendritic cells. These...sharing those ideas with others. This love of interacting with people and talking about science makes her...

...behaviors. Scientists continue to engineer opsins to operate at increasingly faster kinetics. The opsin ChETA...body. Administering the drug once can result in several hours of neuronal excitation or inhibition. This...sustained activation period is beneficial for use in therapeutic or behavioral applications where drawn out effects... by Designer Drugs): Chemogenetic Tools with Therapeutic Utility. Annual Review of Pharmacology and Toxicology...

...of time, especially at room temperature or at 4°C. Storing DNA: temperature and longevity You should consider...molecular biology like restriction enzymes and polymerases. In the situation where you need to use plasmid...Plasmid DNA can be stable at 4°C or even room temperature for a short period, and there are indications...

...blot and immunofluorescence. GST  Glutathione S-Transferase (GST) is a 26 kDa tag which can be isolated with...Purification of His fusion proteins from E. coli is generally more straightforward and of a higher purity than...washing. Nonetheless, this tag can be purified with several different resin types and is a fairly small tag...and other qualities of each tag class, they are generally best suited for their primary application. Ready...

...cell line collections websites and catalogues Literature searches utilizing Google Scholar alerts and ...feature to know if your lines may be especially vulnerable can be very informative. Spoiler alert: HeLa ...information on the known features and relevant literature, or to decide which line is best for your experiments...Josef Reischig.  Contributing to Cellosaurus Generated a new cell? Or finding errors in the available...

...receptor ACVR1. ACVR1 triggers appropriate bone regeneration in healthy patients. However, the R206H mutation...including Activin A. A commercially developed therapeutic antibody that binds Activin A and interferes ...interferes with inappropriate activin-ACVR1 interactions prevents FOP bone growth in mouse models. As this biologic...experience is in both vaccine subunit design and therapeutic biologic design (ADCs) along with pre-clinical...

...escape reflex. These traits, coupled with short generation times and high fecundity, make zebrafish ideal...lights (Baier et al., 2009).  Zebrafish behaviors Several behavioral assays can be done on zebrafish at 5...neurons that trigger the CPG (central pattern generator) (Wyart et al., 2009). The CPG is a neural circuit...circuit involved in locomotion that generates periodic motor commands for rhythmic movements like spontaneous...

...the the University of Cambridge and within the general public. The society organises talks by prominent...graduate students, postdocs, faculty members, and the general public. Chemical Society in University College ...studying to earn a PhD. The society encourages interactions between undergraduates and PhD students so that...develop a range of talks for both students and the general public, career events, and networking meetups. ...

...included in organizational decision-making. It is imperative that employees are thanked for their work by ...but it is worth it. This requires a focus on deliverables. It does not mean a manager must allow his team...enough of a jerk to make the rest of the team miserable.   A good manager keeps these factors in mind...decisions, but that should not be the common mode of operating.  Managing scientists I should add a few special...

... concept that these clustered repeat sequences operated as a safeguard against bacteriophages did not ... bacterial genome. Cas genes, which code for polymerases, nucleases, and helicases, were also disrupted...proteins then express and process the CRISPR loci to generate RNA, which guide a Cas nuclease to the specified...learned to circumvent the CRISPR/Cas defense by generating a single point mutation in the PAM sequence, ...

...study which can be accessed here at bioRxiv. AAV2 interacts with the packaging cell's cell membrane When packaging...blocked following sucrose treatment. Moreover, several reports have shown that cells treated with sucrose...    Despite observing a positive effect on our overall yield, we were unable to demonstrate that these...-associated virus manufacturing." Human gene therapy21.10 (2010): 1251-1257. PMID: 20649475. PubMed Central...