...his own biases--many of which will be shared by fellow scientists--but that doesn’t make his introduction...have outgoing personalities to supplement their intellectual prowess and scientific skills. This is an era...has been such a great fit for me!), the book was a well-executed overview that recontextualized some areas...career.  Acknowledgements: I would like to thank fellow Addgenies Joanne Kamens, Marcy Patrick, Aliyah ...

...using an electric charge to help the DNA enter the cells. Some samples require a reduced antibiotic concentration...concentration or need to be transformed into different cells than were originally requested. Most of our samples... Start by scanning the sample barcodes into the cells in the same order that they exist within the rack...storage process - from deposited DNA to transformed cells and finally to bacterial glycerol stocks that are...

...Many thanks to our guest blogger Gary McDowell! Gary S. McDowell is currently the Executive Director of...by Future of Research Executive Director, Gary McDowell. On Dec 1st, the threshold at which salaried workers...come under this ruling, regardless of visa or fellowship status, in addition to certain staff scientists...

... be safely used to deliver a genetic payload to cells, and the complexity of effectively deploying them... can be to use. AAVs, which can target different cell types depending on their serotype, are particularly...amount of vectors available and number of requests tell only part of the story. Viral vectors are, after...experimental data in a standardized format, sharing how well a given viral vector worked — or didn’t work — in...

...designed to help us work in large formats, such as 96-well microplates, and thanks to our dedicated product...: Customizable to our specific needs Modern and well-supported, so we could focus on our development ... workflow would handle plasmid sequences on a 96-well plate basis, but for rapid testing and refinements...blog:  The Power Behind NGS Plasmid Validation: SeqwellHow to Keep a Lab Notebook for Bioinformatics AnalysesAAV...

...other journals and several funding agencies (e.g. Wellcome Trust, NIH Data Sharing Policies, etc.) and is...numerous objections and concerns. Perhaps least compelling of these were those from researchers who wanted...released, we will be in danger of having to deliver on well-meaning but impractical requirements imposed by ...should open up new opportunities for research as well as increasing transparency. Thank you to our guest...

... The loading control is supposed to control for, well, loading errors. When using it to normalize your...Most housekeeping proteins are highly expressed in cells. This means that at sample concentrations appropriate...assay. This is influenced by sample concentration as well as by antibody dilution — both factors we’ve covered... C, indicating that β-actin is less sensitive as well as less linear than the total protein quantification...

...academia Require that postdoc salaries be paid on fellowships instead of grants such that PIs no longer save...brought in editors from a variety of publications as well as young PIs and all gave advice on how to succeed...Rafael Luna, Joan Reede, Alberto Roca, and Jessica Tytell made it was obvious that more data is needed for...discluded/not treated equitably even when hired; well-meaning mentors often fail to judge the work of ...

...Every time I asked a question, I felt like I was yelling. I imagined 6 years of biting my tongue and gratefully...lab across the hall or in the same department as well. What do they have to say about this lab’s reputation...appointments with group members or are they frequently cancelled? Choosing a role model Make sure the mentors ...target up to fail, shifting goal posts without telling the target Warning signs while interviewing: Lab...

...mismatched) around the ligation junction in 16 wells of a 96-well plate, allowing 6 conditions to be screened... (2006): 687-699. PubMed PMID: 16464020.  3. Ellenberger, Tom, and Alan E. Tomkinson. "Eukaryotic DNA ...ligase bound to nicked DNA-adenylate." Molecular cell26.2 (2007): 257-271. PubMed PMID: 17466627. 25. Luo...

...function using transcriptional recording sentinel cells! 🎵 On the 11th day of CRISPR, Addgene gave to me...Enhanced prime editing systems by manipulating cellular determinants of editing outcomes🎵 On the 5th ...activation and interference screens for primary human T cells🎵 On the 3rd day of CRISPR, Addgene gave to me: ...

...ImagingResearchers: Andrea Radtke, Ziv YanevCellosaurus: A Cell Line Information DatabaseResearcher: Amos...Undergraduate Research Experiences (CUREs)Author: Jason PellettieriNew Acoustic Reporter Genes: Ultrasound Imaging...

...Protein Plasmids 101: Visualizing Subcellular Structures and Organelles Resources at Addgene.org Find ...Plasmids Find Plasmids for FRET Find Plasmids for Subcellular Localization ...

...multiplex gene editing: Guest blogger Qichen Yuan tells us about a new tool for efficient multiplex base...Demethylation: Guest blogger Daniel M. Sapozhnikov tells us how to use dCas9 (with no modifications!) as ...research story, I couldn't resist adding in this excellent Q&A from Tim Miles on everything from AAV prep...

...next week - be sure to come by booth #851 and say hello! Just in time for this meeting, Addgene is launching...synthetic biology, gene cloning and expression, as well as guide pages for Synthetic Biology and Bacterial...of a useful plasmid that’s not in the collection? Tell us about it with our Suggest a Plasmid Form! Finally...

...Mammalian cells provide scientists with the means to study biological processes on the cellular level instead...with tissue culture The cultivation of mammalian cells in the lab, or tissue culture as it is commonly ...

... Immune cells Flow cytometry (potentially others) Data repository From the Human Cell Differentiation...repository   HuBMAP Consortium Healthy human cells Imaging (IHC, ICC, IF, etc.)  Data repository...

...nuclease target multiple genes in a cell population. The resulting mutant cells are screened for phenotypes of...range. CRISPR can be difficult to deliver. Not all cells efficiently take up plasmids used to produce CRISPR...

... serotype pAAV-hDlx-Flex-dTomato-Fishell_7 AAV2 Controls Fishell New serotype pAAV-mDlx-NLS-mRuby2...of GCaMP, GCaMP8, available in new serotypes, as well as seven vectors newly available in the AAV PHP....

...projection neurons and mPFC interneurons can be labelled with an AAV1 Cre virus combined with a Cre-dependent...includes: Images that clearly show viral spread and cellular tropism* (min. 300 dpi, PNG format) At least two...channelrhodopsin Images showing the injection site as well as the target region, if the report describes a ...

...Say hello to the new Addgene blog! Same URL, same great articles to help you with your experiments, but... search, you’ll see the blog articles come up as well as their topic category. New author pages to find...Get in touch by emailing us at [email protected] or tell us in the comments below. We’ll track down the story...

... ranging from new tools you can use to monitor cellular perturbations to how CRISPR has made it much easier...2016 Click on the links below to read all of our excellent posts from July: Addgene at the ASM Microbe 2016...Studies Gendered Innovations: Why Does Sex of the Cell Matter? AAV: A Versatile Viral Tool for Gene Expression...

...more about. Our post on “Making Your Own Competent Cells” has been extremely popular, generating many additional... We’re happy to see scientists making their own cells to save money and create the tools that work best...Save Time and Money by Making Your Own Competent Cells By Melina Fan Plasmids 101: The Promoter Region...

...Multiplex genome engineering demonstrated in human cells They used the Golden Gate cloning method to ligate...successfully to target multiple sites in cultured human cells. While it has not yet been demonstrated, they say...Reference: Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. Sakuma...

...real lab!” And several of our visitors made sure to tell us what we already knew about our Shipping Operations...awesome!”. The students also got to learn about intellectual property from our Tech Transfer team members...After hearing about many Addgenies’ career paths, Michelle Healy said “You are always learning and that means...

...genome-wide deamination of RNA cytosines in human cells. Thus, they engineered two CBE variants with mutations...RNA edits by >390 fold and >3,800 fold in human cells with no significant reduction in editing efficiency...mice and in zygotes. Read the paper in Molecular Cell Find the Nme2Cas9 All-in-One AAV constructs CRISPR...

...doing some research, notably determining the subcellular localization and the number of disulfide bonds...in your protein - two parameters that affect how well E. coli can produce your protein. Structural information...system. If you find that your protein might not fold well, you should screen a few different vectors to see...

...mashed potato bar (complete with all the fixings), as well as plenty of delicious desserts. We enjoyed a video...the changes at Addgene over the last 10 years as well as the many people that make up the Addgene family...Anniversary Gala. The Addgene lab often works as a very well-oiled machine and this post was no exception - the...

...21st century and all. But while the auto-reply telling you your message was received is a nice touch, ...answer your question. Does this sound familiar? Well, when it comes to working with Addgene, we certainly...and requesting them from your colleagues, to intellectual property rights and handling your constructs...

... modified Ti plasmids during plant infection, as well as plasmid yield during preparation in E. coli. ...both β-glucuronidase (GUS) and GFP reporters, as well as multiple transient and transgenic systems, Sahoo...versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants....