... more than 10 modules often rely on two-step hierarchical approaches using different Type IIS restriction...stability, and buffer compatibility have placed practical limits on single- or two-step assemblies. We have...upon growth on LB/Cam/X-gal/IPTG agar plates, indicating successful reconstruction of the coding sequence... While T4 DNA Ligase, the mainstay of most biotechnological cloning efforts for over 50 years, prefers...-Crick base pair substrates, it demonstrates significant activity on some mismatch-containing pairings... sequence-dependent ligation efficiency and identification of mismatch-prone pairings. Using these 4 bp...Kucera is a Principal Development Scientist of Applications & Product Development focusing on DNA cloning...

...example, if you’re a virologist presenting at the American Association for Cancer Research Annual Meeting...therapeutic strategies to target it, than you typically would at a microbiology meeting. Create an outline...lab mates, and PI can provide feedback on your technical presentation, but those outside of your field ...don’t fully understand the question, ask for clarification. Having them reframe or elaborate their thought... through the list at the session so you can strategically use your time. Don’t have access to a list of...titles to find the ones you’d like to revisit. Strategically traverse the sea of posters If you come prepared...system, and is interested in a career in science communication.       Resources on the Addgene blog Read other...

...recognized by its antibody without additional chemical modification steps and when the epitope was within the...antibodies for every potential new target and basically impossible if you wanted to distinguish between...C-terminus post-translationally, which turned out to be critical for antibody recognition. As such, the peptide...interest and proteins tagged with it needed to be chemically treated before the antibody could detect them... the N-terminal half of the antigen differs significantly between human and the other species, suggesting... flexible - it could now be used in way more applications.   Fig. 1 Making a Protein Tag (Abridged... of new versions of Myc1-9E10 mentioned above basically involve taking the Myc1-9E10 variable regions ...

...transformative biological discoveries and life-saving medical breakthroughs and undermine critical research ...Addgene as a trusted partner for archiving their biological materials during funding transitions. Enabling...operating model allows researchers to access authenticated materials at a fraction of the cost of creating...be listed as direct costs from a CRO in grant applications, addressing the new funding structures and indirect...private partners to create additional pathways for critical scientific work to continue.  Moving Forward ...

... citations for recent articles. Of course, a publication in Science or Nature is always nice, but in the... lesser-known journal as it is in a top-tier publication. Accessibility is surely also behind the increase...increase in citations for older publications. (As an interesting aside, Nature recently reported that the...in a solution.) But are citations and journal publications really the best way to measure impact? As that...papers detailing Nobel Prize-winning research typically weren’t among the most cited. And, as scientific...and what it’s made of. Increasingly, journal publications alone aren’t enough. Scientists are sharing ...consider and quantify the impact of scientific publications. Ventures like Impact Story seek to find ways...

... and chemical contaminants that have been used during the purification procedure. Good indicators of DNA... is necessary for bacterial transformation of chemically and electrocompetent cells is minimal (picogram...nm should fall ~1.8 - 2 and give scientists an indication of purity versus protein contaminants. The 260...nm ratios should fall ~2 - 2.2, and provide an indication of purity versus chaotropic agents as guanidine...addition to being free of any nucleic acid and chemical contaminants, these DNA samples must also be entirely...

...immunoprecipitation applications, but none of the traditional tags were suitable for both applications. To prove...from non-vertebrate systems. Preliminary testing indicated that inntags IT5, IT6, and IT10 expressed well...and the chaperone Ydj1, both of which display sophisticated interactions with multiple other proteins. Again...detrimental effects on cell size and growth rate, indicating that they compromised protein-protein interactions...These tags are easy to use in multiple common applications. Due to their limited effects on native protein...particularly in single-molecule or high-throughput applications. Perhaps IT5 and IT6 will even replace our older...

...Balancing research with medical practice as an MD/PhD. Darrel does 20% clinical work 80% research and was...for Regenerative Medicine at Boston University Medical School. Darrell’s research focuses on the lungs...Center for Regenerative Medicine, and within the biomedical research community. Along these lines, Darrell...Pathology and Laboratory Medicine. He attends in the Medical Intensive Care Unit, the Pulmonary Consultation...the Pulmonary outpatient clinic, all at Boston Medical Center. In addition he conducts basic stem cell...Regenerative Medicine of Boston University and Boston Medical Center. Special thanks to our sound engineer/producer...

...conventional microscopes and give a similar optical quality (magnification of 140X and 2 micron resolution). I... science communication and outreach - if you'd like to learn more about Science communication check out...listening to talks all day. But nope, not at the American Society for Microbiology 2018 Microbe meeting....biomolecule-based dyes. This technology is widely applicable as bacterial expression of dyes allows dye visualization...Bacterial Genome Engineering Learn about Science Communication using Twitter  Resources on Addgene.org Check...

...provided here for reference: OpenWetWare Preparing chemically competent cells (protocol)  Preparing electrocompetent...protocol)  Untergasser Lab Preparation of chemically competent cells (protocol) Preparation of electrocompetent...until we are ready to perform transformations. A typical 200mL bacterial culture produces 20mL of competent...transformation protocols below: OpenWetWare Transforming chemically competent cells (protocol)  Electroporation (...protocol)  Untergasser Lab Transformation of chemically competent cells (protocol)  Transformation of...being performed at Addgene, this adds up to a significant time savings. You can also save yourself time...plasmid is introduced to bacteria, the bacteria is typically grown in liquid culture without antibiotics to...