...interfere with it in a BSL-2 lab instead of a BSL-3 lab. Jesse Bloom’s lab published a protocol and reagents... CRISPR-Based Detection Tools. The CRISPR Journal 3:142–145. https://doi.org/10.1089/crispr.2020.0030 ...a method that uses loop-mediated isothermal amplification after reverse transcription (RT-LAMP) and a ...receptor for SARS-CoV. The lab also found that cathepsins play a role in SARS-CoV infection and deposited...deposited plasmids for the expression of cathepsin S, L, and B. This became a great starting point for SARS-CoV..., Zengel JR, Sathyan V, Nguyenla X, Van Dis E, Catamura C, Yamashiro LH,  Begeman A, Stark JC, Shon DJ...

...the gel and add gel components to polymerize. Step 3: digest the sample to allow for even expansion.  ...:832–863. https://doi.org/10.1038/s41596-018-0117-3 Wassie AT, Zhao Y, Boyden ES (2019) Expansion microscopy...with an abundance of protocols and examples of applications from across disciplines now available. In this...experiment or evaluating ExM images in a new publication.         Why Expansion Microscopy?  Traditional...microscopes sounds great, right? But there is always a catch - some limitation of the technique that users should...Tissue expansion is not compatible with in vivo applications and there’s just not much to be done about that...

...pathway (Charpentier et al., Nat Com).     Fig. 3: HDR events compete with NHEJ-mediated edits of small...Non-Homologous End Joining Learn about Base Editing Read 3 Tips to Improve HDR Efficiency for CRISPR Editing ...cut site location Position your CRISPR cut site as close as possible to the knock-in location. HDR depends...sequence to be incorporated at a precise genomic location, with little to no room for error. It’s no surprise...nutrients in their media, they will not undergo DNA replication, so make sure you are keeping an eye on those...

...assessment of many protein targets in a single sample; 3) producing recombinant, partially humanized rabbit...target proteins of interest for labeling, quantification, purification, chromatin immunoprecipitation and more...created the Monoclonal Antibody Research Institute dedicated to: 1) developing new ways of generating antibodies...Freedom: The Expansion of Data Sharing in Research Publications ...

...into Stage 3 or Stage 4 cancer. These later stages of a cancer diagnosis indicate, for Stage 3, that the...large in size and may have spread while Stage 4 indicates that the cancer has spread from where it originated... dire prognosis for mesothelioma patients is to catch the disease earlier. New techniques are currently... working to fight the cancer.  Beyond early modifications to standard treatments and early detection, ...

... for an extended period of time (>10 passages or >3 months). Other times cell lines are unavailable from...State University of New York Press, Albany (1986). 3. Masters, John R. "HeLa cells 50 years on: the good...Marker-independent identification of glioma-initiating cells. The International Cell Line Authentication Committee...propagation of the line, and can include assays for authentication and contamination) and cell lines can become...let alone the human cost. Here are just a few publications which have since been retracted due to cell ...

... kb at most - which means that a gene larger than 3 kb cannot be delivered using this vector. For example...emerges from disgrace to be the next big thing, again 3. Acland G. et al., 2001. Gene therapy restores vision... time advances in human genetics led to the identification of several disease-causing genes, including... But it is the hard work and persistence of a dedicated team of scientists led by Dr. Jean Bennett (UPenn...-1250015778 7. Yu W and Wu Z., 2018. In vivo Applications of CRISPR-Based Genome Editing in the Retina...

... dangle the carrot (or more popularly- cookies). 3. Do something outside of your ordinary environment...Utilize your office space too Balance point number 3 with doing fun things within your office. This way...be an obvious one, but don’t make things too complicated for yourself (especially if you are the sole ...sole event planner)! If you have a very intricate itinerary, there is a higher likelihood that something ...Delegate- it will save you time and sanity!    10. Communicate Make sure people know that there is something...

...Improved Vectors for Gene Delivery. Curr Gene Ther 5(3), 299–310. PubMed PMID: 15975007. PubMed Central PMCID...Concepts in the pathogenesis of rabies. Future Virol 3(5), 481–490. PubMed PMID: 19578477. PubMed Central...viral vectors is serotype-dependent. Gene Ther 20(3), 348–352. PubMed PMID: 22418061. PubMed Central PMCID...to its retrograde axonal transport. J Cell Biol 82(3), 798–810. PubMed PMID: 92475. Taymans, J. M., Vandenberghe...production and purification that yield high titers (see also AAV protocols for production, purification and titration... are replication incompetent and cannot readily be passed on to subsequent cells via replication like ...intracerebral application, which is more invasive and has a much higher risk of complications. References...

... As a small, tight-knit organization we thrive on 3 things: fun, teamwork, and competition. And our Halloween...plasmids! Each chose their favorite plasmid and replicated it. Can you guess each plasmid? Our lab paid...creative hijinks last year. They showed us their dedication to great customer support by dressing up as our...

...classic lab and home editions. We will select up to 3 winners for each edition and the winners will be revealed...create a stack of wrapped gifts using life science catalog pages. Make your WFH space feel a little more festive...

...type and the specific genes cloned into the vector.3  2A Peptides To overcome some of the disadvantages...et al. (J Virol. 1988 Aug;62(8):2636-43.) PubMed. 3. Highly Efficient Multicistronic Lentiviral Vectorswith...desktop. The Plasmids 101 eBook is designed to educate all levels of scientists and plasmid lovers and...

...with a single insertion because it will segregate 3:1 in the T2 generation. You also have no control over...PMID: 17087829. PubMed Central PMCID: PMC1636043. 3. Kyndt, T. et al. (2015) ‘The genome of cultivated...Agrobacterium tDNA insertion likely precipitated domestication of sweet potatoes (Kyndt et al., 2015)! It sounds...

...the impact of research" (2013) Scientific Reports 3, Article number: 1649 doi:10.1038/srep01649     ... 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...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...

... them. For intracellular targets, we usually test 3 different fix/perms (methanol, PFA-saponin, PFA-triton.... The approach we suggest is summarized in Figure 3. The principle is to confirm the antibody can detect...-use of an individual experiment.   Figure 3: Our recommendations for an approach that may be used...procedure enables quantification of antibody performance in major research applications. eLife, 12, RP91645...that the antibody being published doesn’t always indicate the presence of supportive data. To be supportive...that the antigen being detected in the intended application — here, flow cytometry — is the desired target...fixation-permeabilization protocol. Although the expected location of the target is an important factor in deciding...

...controls” category, which includes things like viral vectors that deliver GFP.   Figure 3: A) Cumulative... why they are the most commonly requested (Figure 3). Over 400 biosensors were distributed in 2023 alone...in 2018. By the end of 2023, our catalog had grown to 879 viral catalog items (Figure 1). The service caught...repository. GCaMP, a calcium indicator (biosensor, by Addgene’s categorization) was first developed in 2001....   Figure 2: Numbers of AAV tools, by category, available from Addgene as of 2023. A large... calcium and voltage sensors, are the broadest category of viral vector tools, which may help explain ...capsid of the AAV, many of the vectors in Addgene’s catalog are offered in different serotypes. In fact, the...

...the world — and we really do mean all over (Figure 3). We continue to add new posts every week, and we’...beautifully designed and easy-to-reference eBooks. Figure 3: Global readership of the Addgene blog in 2023. Views...logistical barriers, allowing people to access a wide catalog of plasmids from all over the world. This idea,...into the accessible. Physical access Addgene is located in the USA, but our materials are available to ... items to countries with a distributor. Arrows indicate the year we entered into an agreement with a distributor...accessibility to researchers around the globe. Addgene’s educational resources As Addgene built out our scientific...guides, protocols, and collections of 2023. Educational Resource Top Five by Views Guides CRISPR...

...errors in experimental design and gel quantification. Figure 3: In a perfect world, protein concentration...versus the amount of protein loaded on the gel (n=3). Data is presented as mean ± SEM. Adapted from Analytical...normalization and loading controls needed for relative quantification of a western blot — and why you might want ...accurately compare different lanes? In relative quantification, the POI bands are normalized to the loading...and it’s also problematic for computational quantification. Outside the linear detection range, band intensity...common error associated with Western blotting quantification” (Gilda & Gomes, 2013). To address this source... Gilda & Gomes, 2013. Figure 7: Semi-quantification of liver lysate using different loading controls...

...transfer plasmid with the E1 and E3 (Early 1 and Early 3) genes deleted. These genes’ functions are rescued...Degeneration in Tissue Culture. Proc Soc Exp Biol Med. 84(3), 570–573. https://doi.org/10.3181/00379727-84-20714...and late gene transcription) is demarcated by viral genome replication (Harrach, 2014). Given that the...encapsidate new virions) are not expressed. Note that replication-competent AdV vectors also exist and are used...the final topic, which is that natural AdVs, replication-competent viruses with a recombinant genome, ...Hermiston, T. W. (2003). Multigene expression from a replicating adenovirus using native viral promoters. Mol ...does not require de novo viral gene expression: Implications for gene therapy. Proc Natl Acad Sci U S A, ...

...understand your proposal to give you proper consideration. 3. Present an easy to follow seminar See item 2. Attendees...? How will your approach resolve this problem? (2-3 sentences) What will be the focus of your first... do need multiple first/corresponding author publications in quality journals. It helps to come from the...a job. The currency of scientific success is publications and grants. 2. Write succinct and easy to understand...humbling. My conviction that I had a flair for communication was rapidly dispelled. She would ask me, "Is...succinct. Slide decks leads to the opposite and can suffocate an exciting idea in a swamp of unnecessary details...essential for human health. Failure to do so is implicated in several human diseases. Proper patterning ...