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...third-generation packaging plasmids. For a summarized comparison of second- and third-generation systems, please... cloning methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may...doi.org/10.1016/j.biopha.2020.110276 PMID: 32502836 Johnson, N. M., Alvarado, A. F., Moffatt, T. N., Edavettal...

... James M. Wilson New serotype pENN.AAV.hSyn.Cre.WPRE.hGH AAV2 Recombinase James M. Wilson New serotype...James M. Wilson New serotype pENN.AAV.CamKII 0.4.Cre.SV40 AAVrg Recombinase James M. Wilson New serotype...FLEX-rev:: hM4D-2a-GFP AAV1 Chemogenetics Scott Sternson New viral prep pAAV-CaMKIIa-hM3D(Gq)-mCherry...resources!   This blog post was written by Rachel Leeson and Brian O'Neill.  More resources on the Addgene...

...PMC4246457. 4. Davidson CJ., et al. "Improving the limit of detection for Sanger sequencing: a comparison of methodologies...although this has been improved in some studies) (Davidson et al. 2012, Tsiatis et al. 2010). As the field... approaches can provide the researcher with a reasonable level of certainly that the effects they see ... PhD candidate in the laboratory of Prof Steve Jackson at the Gurdon Institute at the University of Cambridge...Vouillot, Léna, Aurore Thélie, and Nicolas Pollet. "Comparison of T7E1 and surveyor mismatch cleavage assays...PubMed Central. 5. Tsiatis, Athanasios C., et al. "Comparison of Sanger sequencing, pyrosequencing, and melting...

...Non-science favorites  The Mistborn series by Brandon Sanderson "The Mistborn series is the first 3 books of a...funny, bittersweet, and shocking at times."     - Jason Snair, Product Designer Sarum: The Novel of England...skeptical reader of historical fiction (I can personally attest to this!). Following history from the ...participated in a book club before, but I had a few reasons for launching this one. On a purely selfish level...contentious, discussions. I presented the first book, Nicholson Baker’s The Anthologist. The book ended up literally...Prejudice, and some more obscure treasures, like Tove Jansson’s The True Deceiver. There have been books that ...

...such a resource when updates depend on a single person. With thousands of scientists routinely using and...Protein Property Visualization allows for visual comparison of FPs and how they relate to one another. .  ...you or your lab use frequently) for the sake of comparison or ease of retrieval.  These collection pages...of your choosing!), and they can be downloaded in JSON or CSV format to quickly retrieve all of the information...store and represent within-study photobleaching comparisons.  We very much appreciate feedback, so try out...plasmids Find plasmids for FRET Check out the Michael Davidson Fluorescent Protein Collection ...

...be a graduate student, postdoc, professor, etc. Reasons to pursue a visiting role The main motivation to...can’t obtain at your current institute. A common reason is hands-on training in a technique that only a...hard to find, or inaccessible to you for another reason. Visiting would allow you to train and acquire ...data you need on the new instrumentation. Bonus reasons Visiting gives you access to possible new collaborations... willing to host you! If there is someone you personally know or your lab collaborates with that can fulfill...work together to ensure your expectations are reasonable, your reagents are onsite before you need them...

... Connelly JP, Pruett-Miller SM, Blanc V, Davidson NO, Madison BB (2017) A stable but reversible integrated...assaying CRISPR/Cas-stimulated HDR activity) The Madison lab created SRIRACCHA in 2017 to enrich and identify...Wen et al., 2018). SRIRACCHA uses the Piggybac transposon to stably integrate a reporter gene that contains...your cells.  A plasmid that contains Cas9. The Madison lab also created an inducible SRIRACCHA system ... experiment that targeted the gene NFE2L2, the Madison lab found that H2B-GFP expressing cells exhibited...

...If cloning methods had personalities, SLIC (sequence- and ligation-independent cloning) would be a true...also be beneficial. SLIC is most often compared to Gibson assembly, a cloning method also based on homologous... recombination. The higher temperature at which Gibson assembly takes place may limit formation of secondary... of fragments. The major advantage of SLIC over Gibson assembly is cost, as T4 polymerase is much less...less expensive than the enzymes required for Gibson assembly. SLIC is a standardized method for multi-fragment... Addgene Ligation-Independent Cloning Protocol Gibson Assembly Blog Post Choosing a Molecular Cloning...

... think more favorably of scientists they know personally Scientists in your field will know about grants...Job hunting outside of academia is all driven by personal connections What can you do to get the most from...whose company they enjoy on a professional and personal level.  Here are some tips to get out of the rut...free places to order cards. There are a few good reasons to have a card but two of the best are: if you ...with a more general intro until you know if the person you are talking to is really in your direct area...attend. Meet some people online to follow up with in person. Email lab members that have moved on to see if...

... https://doi.org/10.1021/acssynbio.6b00031. Iverson, Sonya V., et al. “CIDAR MoClo: Improved MoClo Assembly...proteins, then the CIDAR Kit is an excellent option (Iverson et al., 2016). CIDAR offers a one-pot reaction ...developers noticed that studies frequently made comparisons between CRISPR components expressed from different...nature of the Fragmid system allows for direct comparisons of CRISPR parts. The Fragmid system was used ...fungi, bacteria, protists, plants, and animals (Chiasson et al., 2019). Many plasmid elements such as promoters...Crossref, https://doi.org/10.1016/j.xgen.2024.100519. Chiasson, David, et al. “A Unified Multi-Kingdom Golden...

...requests without a personal message and they may report you as spam or “I Don’t Know This Person.” If that happens...Contact info" link right below their photo. 2) Personalize all connection requests LinkedIn is a place to... to build relationships. If you don’t add a personal note, you waste an opportunity to start things out...Connect.” If you click this box, it will send an unpersonalized, generic invitation. Don’t try to connect from...have had success with this and as long as you personalize the invitation with your intention for connecting...

...CRISPR-Cas systems.   Four years later, the Sontheimer and Davidson labs discovered three anti-CRISPRs for ...Amrani N, Chen JS, Cofsky JC, Kranzusch PJ, Sontheimer EJ, Davidson AR, Maxwell KL, Doudna JA (2017) A Broad-Spectrum...Hidalgo-Reyes Y, Lee J, Edraki A, Shah M, Sontheimer EJ, Maxwell KL, Davidson AR (2016) Naturally Occurring Off-Switches...

...your next project. Gymnotiformes and Mormyridae (Jason Gallant) Weakly electric fish are excellent systems... 60 °C soils. Dendrobatidae (Lauren O’Connell)  Poison frogs are great for studying parenting strategies...generate cardenolides that tastes yucky (and is poisonous) for bird predators - birds eat a caterpillar ...of a pigmentation gene.   Genus Microtus (Zoe Donaldson) Voles have fascinating social behavior. For example...Claudio Mello) These very cute birds learn their songs like humans learn speech. Finches must learn early...system to rapidly identify recombinants. Arthropods (Jason Rasgon) This lab is developing tools for pan-arthropod...

... make sure to get that person’s contact information and again, make a personal connection! Even if you...does, however, offer you an opportunity to make a personal connection; the importance of which will be elaborated...first for companies at which you’ve already made a personal connection. That way, once you’ve identified an...email with the relevant info and they just might personally hand your credentials to HR, greatly increasing...information and establish a connection with that person before ever asking them for work. Addgene’s very...

...difficult to find solutions to. There are many reasons why client situations might be especially challenging...skills is a must to be successful in this field. Personally, I have greatly benefited from and appreciated...when their teams are facing a crunch, and using lessons from previous cases to better staff future teams...conflicting work styles, team situations, and personalities. Teams striving to achieve a lot of great output...coach and counsel employees on the different personality and work styles (e.g., Myers-Briggs). Staff are...teams, to leverage the preferences of different personality types, and to devise strategies to be prepared...

...BRET= Bioluminescence Resonance Energy Transfer CRET= Chemiluminescence Resonance Energy Transfer    Interesting...now well-known FRET, aka Fluorescence (Förster) Resonance Energy Transfer, technique which allows the detection...have any cross-talk but you also don’t have any resonance transfer. You have to find a balance between FRET...downloads/modules/FRET_teaching_module.pdf Day RN, Davidson MW. Fluorescent proteins for FRET microscopy: ....201100098. Sekar RB, Periasamy A. Fluorescence resonance energy transfer (FRET) microscopy imaging of live...

...the question of how much data was necessary and reasonable. PLOS has since clarified their data sharing ... in having exclusive access to data and enjoy reasonable time to make sense of it, with their decision...The real cost of making data accessible A more reasonable concern with the data policy was raised by researchers...how results were obtained and this alone is a reasonable motivation for increasing both data accessibility...published data. The importance of data sharing The reason this topic is important to my lab is three-fold... welcomed. However, expectations must also be reasonable, sensitive to the data type and made as facile...

...with Gibson assembly. While this is a comprehensive mutagenesis method, a drawback to using Gibson assembly...plasmids ready for deep mutational scanning.     3 reasons to be interested in deep mutational scanning Hundreds...rate of mutagenesis, and low throughput.       Gibson Assembly: An easy-to-use technique that joins homologous...especially single basepair deletions. Therefore, Gibson assembly is better suited for creating small, focused... PubMed PMID: 27723752.  4. Fowler, Douglas M., Jason J. Stephany, and Stanley Fields. “Measuring the ...

...2015b) under a CC-BY 4.0 license. Long et al., Nelson et al., and Tabebordbar et al. each chose to try.... used SpCas9 in conjunction with AAV9, whereas Nelson et al. and Tabebordbar et al. used the shorter ...not subject to nonsense-mediated decay. Although Nelson et al. observed only 2% genome editing in one experiment...persist long-term. In accordance with this result, Nelson et al. find that dystrophin restoration is maintained... E, Bhattacharyya S, Shelton JM, Bassel-Duby R, Olson EN. Postnatal genome editing partially restores ...Dec 31. pii: aad5725. PubMed PMID: 26721683. 3. Nelson CE, Hakim CH, Ousterout DG, Thakore PI, Moreb EA...

...path. Because there is no demand for a favor, the person you ask will be more likely to say yes and more... Informational Interviews are best conducted in person. Perhaps you can offer to buy coffee or lunch at... your interviewee? It is always best to meet in person to develop relationships, but if it is not possible.... Keep in mind that this is a good time to ask personal questions about salary, opportunities for advancement...advancement, challenges and disappointments. These more personal questions can elicit the most valuable insights...