...with practice. You can bootstrap your science communication skills with training programs, which are increasingly... by collaborating as they craft sentences for abstracts and presentation slides, learn from audiences ... draw and the perspective I want them to form? Training in science communication should prepare you to... of charge to accepted students, and we pay for travel expenses to our flagship workshop. In our view,...with ComSciCon, find us on Twitter, Facebook, Instragram, or send an email to [email protected]. Many...strength of story telling Science communication: 9 strategies to get your foot in the door Resources on Addgene.org...

...every diagram portraying some aspect of eukaryotic DNA would become busy and potentially detract from the ...times have you looked at a diagram depicting transcription, or DNA repair, or replication, or any number...dimensional, as you can see by this post’s chromatin illustrations from Leah Bury of Microscopic Art. I started...researching enhancer elements that control the transcription of a gene a megabase away, or looking at the...the histone methylation status at a double-strand break that needs repairing. They may be looking at chromatin... protein ZMYND8 in chromatin recognition of transcription-associated DNA damage that promotes homologous...

...consider spectral overlap. Fluorophores signal are actually quite broad, resulting in spectral overlap,...This information is important, and you’ll need to track it throughout your experiments.  Now that that’s...inhibits HRP. In those cases, thimerosal at a concentration of 0.01% w/v can be used as an alternative antimicrobial...you’ll want to pick fluorophores with the least spectral overlap between them whenever possible. This will...Fluorochromes-Brightness-Chart.pdf Syracuse Flow Core. Syracuse flow core spectral overlap. Syracuse University Flow Core Facility...January 31, 2022, from https://flowcore.syr.edu/help/spectral-overlap/   More resources on Addgene’s blog ...

...initiate a cascade that helps attack extracellular pathogens.  Transportation of antibodies to places they can...new Isotype trading cards!     References and resources References Janeway CA Jr, Travers P, Walport...region, can be bound by special receptors that transport antibodies through cells and into different body... binding capacity or valency of 10 antigens. An extra constant domain replaces the hinge region in IgM...www.ncbi.nlm.nih.gov/books/NBK27106/  Janeway CA Jr, Travers P, Walport M, et al. Immunobiology: The Immune ...

... 49,000 times! Their long-lasting popularity illustrates the utility these tools have in many different...pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP (PX458). lentiCRISPR...lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang lab is used to deliver... David Root lab is a 3rd generation lentiviral transfer plasmid for cloning and expressing new shRNA sequences...the necessary elements for packaging, reverse transcription, and integration needed for the production of... of shRNA in target cells by the addition of tetracycline/doxycycline to the grown media resulting in ...changed to clarify that it does not contain the trademarked Tet-On(R) sold by Clontech. For additional information...

...proper training and guidance to the people working in the lab facility. This can be done by training eligible...and streptomycin. Mycoplasma can pass through filtration methods because of its ability to change shape...culture through various sources that are difficult to trace. These include the laboratory personnel, the serum...can spread through this form of powdered media. Filtration does not ensure complete sterilization as mycoplasma...mycoplasmas, contamination by other bacteria or extracellular fluorescent signals produced by these bacteria...rather than using 0.2-micron filters, for media filtration. There is no 100% assurance that this process...fluoroquinolone ciprofloxacin, ciprobay, zagam, baytril, tetracycline, etc. are available for mycoplasma removal from...

... production and storage, we grow a little extra and transfer 100 µl of glycerol stock to each of 2 96-...through our deposit process, and then we carefully track samples through our lab using barcoded tubes and...such as special growth instructions (temperature, strain, media supplements), suggested sequencing primers...plasmid sequencing If you’re like us, you need a strategy for processing several sequencing reactions in... have different requirements for volume and concentration, and not all providers are equipped to perform...Your Plasmid Learn How Addgene Uses Barcodes to Track Samples Resources on the Addgene Website View ...

...assays such as mRNA data from transcriptomics; independent antibody strategies that compare the staining ...antibody that is directly conjugated to a detection substrate, such as a fluorophore, or a conjugated secondary...antibody validation.  The 5 pillars include genetic strategies that test expression in knock-out or knock-down...knock-down down tissues and cell lines; orthogonal strategies that compare antibody assay results with those...antibodies against the same target; tagged protein strategies that express a tagged version of the antigen ...antibody; and immunoprecipitation-mass spectrometry strategies that isolate an antibody’s immune complexes and...

...milestones in the field. Research strategy The Research Strategy highlights i) the significance and ...weeks before the RFA due date in order for the administrative side of the grant to be established. Take time... information, selection criteria, and award administration information. Carefully read over all parts ...the best approach to the research challenge. Demonstrating expertise in the field and past successes boost...protocol, and offers alternative solutions to demonstrate thorough thought planning on the matter. The ...delegate funds to supplies, personnel, equipment, travel, etc. As a first time budget writer, you may find...

...later used by another team of researchers for neutralization assays, part of a study that examined the repertoire...effective sera from vaccinated individuals are at neutralizing the different SARS-CoV-2 spike variants. You...loop-mediated isothermal amplification after reverse transcription (RT-LAMP) and a colorimetric readout that can... 45 minutes. The team deposited the reverse transcriptase, protease, and Bst polymerase constructs from...toolkit include ones for expressing MMLV reverse transcriptase and Taq polymerase that can be used in a one-step...Particles with SARS-CoV-2 Spike Protein for Neutralization Assays. Viruses 12:513. https://doi.org/10.3390...10.1126/science.abd0826 Tong P, Gautam A, Windsor I, Travers M, Chen Y, Garcia N, Whiteman MB, McKay LGA, Lelis...

...presentation as an opportunity to demonstrate technical and transferable skills that make you qualified ...position, I just had a phone interview. I also transitioned between different lab groups without having ...stories as the characteristic which the story demonstrates I possess.   Bullet points on resumes are like...spark a more natural conversation while also demonstrating that you did your homework before the interview... to highlight your prior accomplishments and demonstrate your science communication skills. Know your ...long! Mine lasted 4.5 hours, which was a lot of extraversion for me. Make sure to get enough sleep the night...

...should acquire during their PhD and postdoctoral training. Addgene executive director Joanne Kamens has ...scientific society in your field. My leadership training began with the NPA where I gained invaluable experience...issues. I applied and was successfully granted a travel award to attend the annual NPA meeting to learn...formats but I learnt to overcome the personal frustration and submit reports in the format requested which...leadership institute’ weekends would entail intense training to develop our leadership and management skills...to think about their career options, gaining transferable skills such as leadership skills can only assist...engaged once you are in a leadership role or in a track towards one.  Many thanks to our guest blogger Juliet...

...xCas9 for transcriptional regulation and base editing Hu et al. also engineered transcriptional activators...activating transcription. Sites containing NG, GAA, or GAT PAMs averaged 56-91% transcriptional activation...subunit ω. This construct is designed to activate transcription from the accessory plasmid (AP) encoding phage...could recognize a library PAM, it would induce transcription of gene III, permitting phage propagation and...pleasant surprise that selecting for one beneficial trait, PAM compatibility, also produced an enzyme with...

...One of the most powerful strategies to investigate a gene's function is to inactivate, or "knockout",...these techniques in mind when choosing a knockout strategy! Knockout plasmids Homologous recombination is... mechanism to accurately repair harmful double stranded breaks, in which nucleotide sequences are exchanged...endogenous promoter, you can use expression GFP to track cells participating in development or other physiopathological...very low, ranging from undetectable to 0.1%. In contrast, new genome editing techniques such as CRISPR ...created in 1989, less than thirty years ago. As traditional knockout and new CRISPR tools continue to be ...Cell Biol. 19: 1217. PubMed PMID: 19731224 PubMed Central PMCID: PMC2782548 Additional Resources on the Addgene...

...variable regions that bind to specific antigens. In contrast, nanobodies are a single heavy domain fragment...involves using a commercial kit for in-vitro translation of the mRNA libraries. Each mRNA molecule in ... forming a ribosome complex consisting of the translated protein (sybody), ribosome, and mRNA. During ...enriched pool of sybody candidates are then reverse transcribed into cDNA fragments that are efficiently subcloned...response to the COVID-19 pandemic, the Seeger lab demonstrated they could use the protocol to rapidly identify...McLellan JS (2020) Structural Basis for Potent Neutralization of Betacoronaviruses by Single-Domain Camelid...single domain antibodies for the conformational trapping of membrane proteins. eLife 7: . https://doi.org...

... non-specific endonuclease (Cas9) and a single stranded guide RNA (gRNA). The ~20 nucleotide targeting...of wild-type Cas9 and a gRNA generates a double-strand break in the target DNA, which, when repaired through...heterogeneous population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within.... Pooled gRNAs are then cloned into lentiviral transfer vectors to create the CRISPR library. CRISPR libraries...most cases, CRISPR libraries are provided at a concentration that is too low for experimental use. Thus, ... first step is to amplify your library to a concentration that is sufficient to generate lentivirus. Be... you can skip the steps above! Cells are then transduced with lentivirus containing the CRISPR library...

...expressing neurons and GFP (green) to label neurons transduced by each serotype. Working at Addgene has ...provide an avenue to more easily solve these frustrating and often months-long quests to find the right... labeled high- and low-magnification images demonstrating vector performance) will be assigned citable...scientists to evaluate how small changes in vector administration can influence AAV expression properties.  By...providing DOIs, this information can be cited, tracked, and appropriately credited, broadening the knowledge...S., & Klein, R. L. (2018). More expansive gene transfer to the rat CNS: AAV PHP.EB vector dose–response...Kalwani, L., Sierra-Mercado, D., Robinson, J. O., Outram, S., Koenig, B. A., Pereira, S., McGuire, A. L....

...data for our DNA-based reagents. Not only do we extract as much information as possible from depositing...automating the bioinformatics steps needed to transform the raw sequencing data into a complete plasmid... to find a suitable software tool that could orchestrate the steps into a continuous pipeline. We also...appropriate place in our workflow without having to translate or convert programs to a uniform language.  The...concepts and vocabulary of Nextflow so that we could translate our pipeline steps into processes and then connect.... After this validation check, we repeated our strategy and copied a few lines of our existing script ...Bioinformatics AnalysesAAV Vector Quality Control: Going the Extra Mile with NGS...

... but just as potent and specific? The two AAV strategies described above showed successful target modification...prime editors. While using AAV to deliver large transgenes might seem daunting, it’s actually a challenge...split AAVs. In general split AAVs break a large transgene in two pieces and package each piece into an individual... individual AAV. When a cell is transduced by both AAVs, the full length gene and/or protein is reconstituted...when at the ends of two proteins, mediates protein trans-splicing, similar to an intron in pre-mRNA splicing...DE, Brown MT, Deshmukh H, Kemp ML, Bao G (2015) Trans-spliced Cas9 allows cleavage of HBB and CCR5 genes...2020) CRISPR-Based Therapeutic Genome Editing: Strategies and In Vivo Delivery by AAV Vectors. Cell 181...

...to be translated into Modern Greek to be easily accessible today. Roughly speaking, we translated our software...Addgenies, development includes everything from tracking plasmids from the moment a deposit kit is sent...Another advantage of Scrum is that it provides transparency to everyone involved about the status of each...details, step-by-step instructions on how to demonstrate the feature. For example, we recently reduced... Most of the time, updating these libraries is straight forward. However, there are times where a library...accelerating research and discovery. We work with transparency, gladly share our knowledge, and love to collaborate...