...chloramphenicol resistance pCRISPR: carries a spacer targeting the gene of interest and kanamycin resistance E. ...strand, among other parameters. .  Stanley Qi lab's CRISPRi plasmids Stanley Qi’s lab demonstrated that a minimal... specific polymorphisms, including antibiotic resistance genes. This approach successfully targeted specific...

... locus, scientists can mutate the transcription start site, splice sites, exons, and/or promoters to knock... of exons. Disrupting promoter or transcription start site function would also help to reveal the impacts...polyadenylation signal downstream of the transcription start site will terminate transcription of most of the...based approaches to study ncRNAs. (A) Transcription start site (TSS) deletion. (B) Mutations of splice sites...

...in helping us understand the nature of your concern. Please be courteous. We understand how exasperating...ready. I've attempted to answer several emails that start: “Hi! We ordered something from you and it’s wrong...last digit: An answer should come soon, but not instantly. Addgene tries to get back to customers within...

...AdV plasmid construct from step #1 and allowed to stay in culture for up to 20 additional days at which...is required if rAdV is to be used in vivo. The standard method for purification of rAdV uses cesium chloride...viral DNA packaged in virions is determined by a standard curve of known quantity and primers specific for...with serial dilutions of rAdV, fixed, and then stained 10 days later. Plaques are counted and titer is...

...review analyzing and explaining the antibody crisis states that up to 50% of all commercially available antibodies... to have an intuitive guide for checking their staining and neuroanatomy without the fear of referencing...please contact us here! Ava Shipman is a research assistant in lab of Zhuhao Wu at the Icahn School of Medicine...Bradbury, A., Plückthun, A. Reproducibility: Standardize antibodies used in research. Nature 518, 27–29...

...widely used base editing systems, ABE7.10 (and the starting point for a new generation of ABEs), consists ... needed to produce 3 concurrent base edits to withstand antibiotic selection. As a result, 8 mutations... which resulted in variants named “ABE8.x”: “8” stands for the eighth round of ABE evolution and the “...frequency over ABE7.10. The ability to use non-standard Cas9 in the ABE8 complexes enables a broader targeting...

...scientists have developed calcium sensors to better understand how this metal ion is trafficked and regulated...selecting a sensor to use for your studies, you can start by asking ‘What is the biological phenomena I want... sensor expression if you aren’t working with a stably integrated biosensor. Ratiometric measurements ...switches from green fluorescence to a red fluorescent state only by simultaneous (1) user-controlled illumination...

...fidelity. NEBridge GetSet Need a set of overhangs, stat? GetSet can do that for you, complete with predicted... °C cycling 4-base Bsal-HFv2® 37 °C Static 4-base Bsal-HFv2, 1x NEBridge Ligase MM ...They first compared 10-piece linear (the native state of the T7 genome) and circular assemblies. While... complex Golden Gate Assembly, a great place to start is the 24-piece lac operon assembly deposited by...

...that might serve as a starting point for developing useful tools. A lucky start As Andrew and Maria explained...questions alone. They readily admit they don’t fully understand the physical mechanism, though they have collected...luck) to get here, he believes MagLOV is just a starting point for much more. If you’re interested in working...

...cell, organelle, and protein localization. For instance, you might dye your cells and look at them under...researchers can look at a single image of a cell and understand how protein localization changes over time.Hallmarks...can be used in any situation where one wants to understand the relationship between the age of a cell, protein...biological event (trafficking to a subcellular location, start of gene expression, development of a cell structure...

...days of structural biology, Byron Rubin, a crystallographer working with Jane Richardson, provided an ... model of the protein in your hand – you will understand the power of a physical model. Until then, you...turn it over and over in their hands. They then start talking to themselves as they compare the features...Under the guidance of a trained educator, these instances of focused attention become magical teaching moments...

...crucial background. Practice a few times while standing at your poster – it’s a different experience to...your computer screen than it is to pretend you’re standing at the session. Present with confidence Now that... asked a question that we don’t fully hear or understand what we are being asked. I am personally guilty...one. Take a deep breath, and if you don’t fully understand the question, ask for clarification. Having them...

...in vivo studies, our scientists perform all the standard quality control assays for AAVs. These steps include... Unfortunately, humans have been known to make mistakes. What if a tube is mislabeled or someone grabs...vectors? Viral genome sequencing Thanks to the establishment of in-house sequencing capabilities and Seqwell...sample at 5%. Detecting 5% contamination is a good start, but to determine the sensitivity of our method ...

...a physiological effect of SARS-CoV-2 toward destabilizing one’s natural immune response to Adenovirus ...Adenoviral vectors are commonly derived from the mastadenovirus members (mammalian-infecting members) of the...delivery and for oncolytic virus in addition to standard uses in gene therapy and in basic research, like...death protein (E3-11.6K) is required at very late stages of infection for efficient cell lysis and release...

...curriculum. It’s through the PTA that I recently started coordinating the annual Math & Science Night, a...scientist. I gave a timeline of my science life, starting back to Mr. Kelley, my 7th grade biology teacher...them had imagined for a scientist. I think that establishing this history helped demonstrate that a) I was...curriculum and offered engaging examples of how understanding genetics is so timely.” Genome engineering, ...

...Our story today starts with a bit of Addgene history... As you may be aware, Melina Fan, her brother ...buidling - they relieved some of the pressure of start up life and maintained their sanity by throwing ... at Executive Director Joanne Kamen’s home- a testament to the familial aspect of Addgene. As our size...will avoid chaos and delays in getting the fun started. I’m aware that  “organizing the fun” sounds a ...

...protocol. Many scientists, publishers, and small start-ups are turning to software apps to solve our need...are just a few promising options to get your lab started:   7 ways to share your lab protocols online Addgene...Check out all the benefits and how you can get started on our deposit page. Protocols.io Website: http...satisfaction of checking off each step as you painstakingly complete your experiment – one small victory...

... the lab somewhat mirrors the complexity of understanding their wild counterparts. Those familiar with...process that isn’t as simple as it seems. Take, for instance, the GCaMP-encoding viral vectors available in...revamped our viral vector selection process to standardize our feedback and evaluation of viral vectors....requestors can upload their experimental data in a standardized format, sharing how well a given viral vector...

...similar or better efficiency than PEmax, the current state-of-the-art. PE6b was very efficient for short, unstructured...fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged iPAK4 from different...distribution and has been verified to perform well in standard IHC and western blot protocols. You can find more... Figure 5: Delipidated mouse brain tissue was stained with Anti-PSD-95 [K28/43R] (70 µm mounted sagittal...

...purpuratus, the spiny purple sea urchin. It’s not a standard model organism, but it was easy to harvest eggs...this wonder of the organism.   It’s not an overstatement to say that CRISPR/Cas9 editing is one of the...which these methods are presently unavailable.” As stated by one of the NSF program directors, this program...Now what? What would you study first? They are starting by seeing if they can change long legged lizards...