...transduced cells and loss of transgene expression (Interestingly, the strong immunogenicity of AdVs is what makes...virion is a non-enveloped particle of ~100 nm, consisting of a capsid surrounding an inner core which contains...use of this system. New and improved systems are still being developed. A detailed protocol for production...mL of sample. The purity of the sample can be estimated by looking at the ration of DNA (260nm) versus...

...and overlapping complexes that have distinct functions. Many existing methods for identifying PPIs, such...additional interacting factors. Problems with Existing Methods for Studying Protein-Protein Interactions... biotinylated. This makes it’s impossible to distinguish proteins that directly interact with the bait...

...glycerol stock. Choose from: ampicillin, apramycin, blasticidin, bleocin (zeocin), chloramphenicol, erythromicin...box for the “Purpose” field. If you ever have a question about what should go into a cell, hover over the...Distribute pending QC. The name of the Principal Investigator and Organization where your plasmids were first...in March 2017 and updated by Angela Holmes and Christina Mork in February 2024.    Additional Resources...

...and application should be considered The first question you may have is whether a monoclonal or polyclonal... 1998). Consequently, if you find yourself in a sticky species-on-species situation, and other antibody...that users choose antibodies that are easily distinguishable from each other. In a direct approach, that... species or isotype that can be probed with a distinctly conjugated secondary antibody. Consider too that...

...pattern of the antibody binding “distinct imaged pattern.” The distinct imaged pattern should align with...comparison on the antibody signal’s location. If a distinct imaged pattern does NOT align with the atlases...through their own analysis of the background noise, distinct imaged pattern, and exposure times. To test the...

...plasmids can lead to far reaching consequences, from wasting time and money on optimizing experiments, to false...all incoming plasmids. If you are interested in requesting one of these plasmids, you can request that it...reference is given, de novo assembly in Geneious can still provide additional insights to resolve the assembly...team leaves a note on the plasmid page to make requesting scientists aware of the identified discrepancy...

...changeover had no impact on our ability to provide requesting scientists high quality data! Addgene's new ... example, the CAG promoter, a hybrid promoter consisting of a CMV enhancer, chicken beta actin promoter... that is useful, but not 100% complete, we will still make this data available as a partial sequence with...develop a PCR strategy suitable for the library in question. We will order primers that can amplify the insert...

... to modify chromatin. A fluorescent protein can still be included for visualization purposes, but the ...flexibility of the single-pot system will allow investigators to rapidly create multiple synthetic chromatin-modifying...CRISPR plasmids for Borrelia burgdorferi from Christine Jacobs-Wagner’s lab produce different levels of...pooled library from Keith Joung’s and Benjamin Kleinstiver’s labs are now available. These libraries encode...

...have specific web sites for postdoc listings, so the method of posting varies between companies. Like most...that they were having a positive experience, I was still incredibly nervous about leaving academia. A lot...seen listed as a requirement on some industry job postings. Also, if you work in industry for any length ...

...disadvantages. Here is a table listing some of them: Characteristic ESC ASC Self-renewal High Low...promising therapeutic tools. The two unique characteristics that define stem cells are: Their ability... For many years, ESCs or ASCs were the only realistic options for scientists working in regenerative...

...Restriction digest screening for ITR deletion is still necessary. To save time and money, I like to miniprep...Production protocol, I take a lazier approach for harvesting my AAV. I skip the PEG precipitation of the HEK293...PEG solution. PEG takes several hours on a heated stir plate to dissolve and requires monitoring so that...autoclaving, which also takes time. Rather than investing all of this effort in preparing reagents, I choose...

...impressive, full-color images of proteins. But they are still flat. To truly appreciate the 3D structure of these...models function as thinking tools – eliciting questions like “what does this model representt?" ... "how...creating physical models of proteins that are being investigated in their labs. Soon after we began working with...of the night dreaming about the conformational gymnastics that this protein must undergo as it interrogates...

...important to isolate females from males while they are still virgins. Females begin mating ~6 hours after hatching...the Classical Genetics Simulator website. One interesting thing to know is that males do not recombine ... of Fly Pushing by Ralph J. Greenspan. It’s a fantastic book for newbies on how to work with flies. ... convince him to use Twitter @jonchowphd but he still reads “#” as a “pound symbol”. Additional Resources...

...as a creative problem-solving exercise? Asking questions like the below can help you understand the perspective... related to the actual data collection task but still be beneficial to both.  Coming up with creative ...with the company so they know I’m excited while still being able to take my vacation? I want to learn ... How can I negotiate to show the company that investing in me is also beneficial to them? I have an idea...

...within a multi-tRNA synthetase complex, the team investigated whether the generated antibodies could bind ...myo-inositol oxygenase), providing enzyme protection while still allowing substrates and products to diffuse freely...encapsulated enzyme produced 20% more D-glucaric acid, suggesting that MPyV system can improve metabolic pathway... a ready-to-use viral prep in serotype AAV1. Distinguish dendritic neurites from axonal neurites with ...

...kinds of CRISPR tools people were depositing and requesting each year (Figure 2). We did a similar analysis...2012 to 2023; data are smoothed using locally estimated scatterplot smoothing. (C) Number of requests ...continue to develop and share new tools, while our requesting labs continue to request and use new tools. Because... each country. Countries shown are the top 10 requesting countries. FR=France; KR=South Korea; CH=Switzerland...

... the “Align two or more sequences” option to drastically reduce waiting time for search results. Optimizing...with the "Align two or more sequences" option and pasting your expected protein sequence into the Subject...webservices/#pairwise_sequence_alignment_psa Happy BLASTing! Do you have any tips for using BLAST to confirm...plasmid sequencing results or comments on our suggestions? Share your thoughts here to help other labs ...

...is currently investigating the molecular biology of synaptic transmission and plasticity.     Javier García-Nafría...co-transformation with linearized vector backbone. DpnI digestion - 1 μl DpnI is added to the PCR mix to remove ... García-Nafría is an investigator scientist at the MRC Laboratory of Molecular Biology. He is interested...

...-wide screens to effectively and efficiently investigate gene function. CRISPR libraries can be used to...labs of David Root and John Doench at the Broad Institute have developed human genome-wide CRISPRko (knock...lowly expressed genes or genes that are more drastically affected by gene activation rather than depletion...resistance in A375 cells. The SAM system (CRISPR/Cas9 Synergistic Activation Mediator) uses an improved activator...

...editor, the lab started with the Cas13 scaffold, testing a whopping 21 Cas13a orthologs, 15 Cas13b orthologs... the plasmids for the RESCUE RNA editors here! Testing and improving RNA editing from the Cas13b REPAIR...knockdown is more precise than shRNA knockdown, it still displays substantial off-target activity. By mutating...Addgene. East-Seletsky, Alexandra, et al. "Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA...