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TypeBlog PostPublishedMay 2, 2019, 12:41 p.m....share all of these new organisms continues to be challenging. Organism specific viral vector systems would...thus making progress on the NSF “G to P grand challenge.” Creating a productive, new scientific community...So, think beyond Drosophila melanogaster or Saccharomyces cerevisiae (both fine research organisms) - ...social bonds that they form including genetic mechanisms that control who a female wants to mate with ... first? They are starting by seeing if they can change long legged lizards into short legged lizards. ...
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Lab to Office Culture Shock
TypeBlog PostPublishedFeb. 5, 2019, 1:04 p.m....counter-intuitive, but I find office work more mentally challenging than lab work at times.” Sometimes a desk job...experience will be a little different. Some of these changes you may enjoy immediately and some of them may ... just take some getting used to. Just remember: change is good. You can experience a real onboarding...to non-scientists can be a fun and gratifying challenge. You’re also getting the opportunity to learn ...eventually. If you approach that transition dreading the change, the adjustment period will be that much longer... -
New CRISPR Tools: Cas7-11 and PASTE
TypeBlog PostPublishedFeb. 17, 2022, 2 p.m....and Discovery As part of their quest to find, characterize, and develop new CRISPR systems and technologies...proteins that could cut RNA or DNA via a CRISPR mechanism. The weird black pellet's DNA yielded a new CRISPR...Prime editing — the ability to now make any small changes, including base edits, insertions, and deletions...right integrase for this new approach proved a challenge. The old standby of CRE/flp didn’t work – it turns...be independent of gene repair.) This lowers the chance of the insert being “dropped” in the wrong place... -
AAV Titers: Where Do They Come From And What Do They Mean?
TypeBlog PostPublishedNov. 15, 2017, 3 p.m....methods measure different characteristics of the solution. These characteristics give us information on ...temperature parameters. For example, we found that changing the annealing temperature of the PCR from 60° ...al. "Systemic errors in quantitative polymerase chain reaction titration of self-complementary adeno-associated...PMCID: PMC3216358. 4. Lock, Martin, et al. "Characterization of a recombinant adeno-associated virus type... -
Transferable Skills Guide: Career Planning Resources
TypeBlog PostPublishedNov. 14, 2017, 2 p.m....better prepare for your career, and increase your chances of landing that next coveted job! Professional...development (or lack thereof) One of the biggest challenges facing students today is that many graduate and...your credentials to HR, greatly increasing your chances of getting a phone interview! If you don’t have...touch and reach back out when you are looking to change positions in the future. Anyone looking for work... Michael G. Lemieux... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog PostPublishedOct. 19, 2017, 1:54 p.m....inverted vectors expressing light inducible ion channels (plasmid # 20298 for example) have revolutionized...FLEx vectors expressing the light activatable ion channel under the control of Cre recombinase are used in... M. Sternson. 2008. “A FLEX Switch Targets Channelrhodopsin-2 to Multiple Cell Types for Imaging and Long-Range...Sadowski. 1985. “The FLP Recombinase of the Saccharomyces Cerevisiae 2 Micron Plasmid Attaches Covalently...PMC41617. 7. Metzger, D, J Clifford, H Chiba, and P Chambon. 1995. “Conditional Site-Specific Recombination... -
Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs
TypeBlog PostPublishedOct. 27, 2015, 3 p.m....to interfere with the function of resident ER chaperones and may impact cellular function or possibly ...For example, the formation of inappropriate inter-chain disulfide bonds between FPs blocks Golgi complex-localized...protein activity. We encourage investigators to characterize FP-fusion proteins functionally relative to ...moxFPs. The inherent brightness and spectral characteristics of the moxFPs are comparable to the non-optimized...postdoctoral fellow at the University of North Carolina at Chapel Hill. Under the co-mentorship Blossom Damania and... -
Hot Biosensors 2022: Year-End Roundup
TypeBlog PostPublishedDec. 6, 2022, 2:15 p.m....sensor relies on a change in localization to the cell membrane and not a direct change in the fluorescent...better response kinetics and photochemical characteristics than previous NIR calcium reporters iGECI and... Matlashov ME, et al. Design and Initial Characterization of a Small Near-Infrared Fluorescent Calcium... B) LiLac responses are highly resistant to pH changes. Image adapted from Koveal et al. 2022, under CC-BY... -
Filming Science Videos in the Age of Social Distancing
TypeBlog PostPublishedMay 20, 2021, 1:15 p.m....found it easy to drop into a chair and chat with me remotely via an audio chat (Fig. 4). Learn: This works...small operation, so sometimes it can be a big challenge. But ask any filmmaker or videographer and they...you can usually see a director sitting in their chair not watching their actors live on set, but peering... -
Multiple Plasmids at a Low Price: Kits from Addgene
TypeBlog PostPublishedSept. 14, 2016, 2:30 p.m....centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013. These terminators can...: 2388-2393. Huang, Shijie, Xuechen Zhu, and Charles E. Melançon III. "Detection and Quantification...metalloproteinase: López-Pelegrín, Mar, et al. "A novel mechanism of latency in matrix metalloproteinases." Journal...size from 4 to 156 kDa, enabling the user to characterise the efficiency of their cell-free system in ...5373-5383. Walkiewicz, Katarzyna, et al. "Small changes in enzyme function can lead to surprisingly large... -
Introducing the Addgene Antibodies 101 eBook!
TypeBlog PostPublishedOct. 29, 2024, 1:15 p.m....designed to be a helpful desktop reference. Each chapter brings together all of our 101 blog posts on an...to antibodies as research tools, you may find “Chapter 1: What is an Antibody” quite useful. If you’re... design your first flow cytometry experiment, “Chapter 7: Sorting” is there to help guide you through ...find an antibody that works, we highly recommend “Chapter 3: Finding the Right Antibody for Your Experiment... -
A Control for All Seasons
TypeBlog PostPublishedOct. 5, 2023, 1:15 p.m....positively charged groups, such as aldehydes in fixed samples. To address this, block positively charged groups...the label control for all new samples, protocol changes, or when high background labeling occurs (Burry...critical for data interpretation, especially when changes in protein expression are the experimental readout... References and resources References Burry, Richard W. “Controls for Immunocytochemistry.” Journal ... -
Viral Vectors 101: AAV Variables That Matter
TypeBlog PostPublishedJune 1, 2023, 1:15 p.m....promoter, and titer. Figure 1: Flow chart depicting a) steps for selecting serotypes and promoters... different receptors, co-receptors, and other mechanisms to gain entry into the cell (Dudek et al., 2018... different brain regions. Image modified from Aschauer et al., 2013, Figure 1. Used under Creative Commons...the region you want to cover - and as your titer changes, so may your needed volume (de Backer et al., 2010....2014.00076 References and Resources References Aschauer, D. F., Kreuz, S., & Rumpel, S. (2013). Analysis... -
Plasmids 101: Repressible Promoters
TypeBlog PostPublishedDec. 20, 2022, 2:15 p.m....added, it binds tTA, resulting in a confirmation change that prevents binding to the promoter and turning...added, it binds cTA, resulting in a confirmation change that prevents binding to the promoter and turning...this way, you can interrogate the activity of uncharacterized promoters. Similarly, placing UAS upstream ...1472-6750-6-43Kaczmarczyk A, Vorholt JA, Francez-Charlot A. Cumate-inducible gene expression system for ...genes for metabolic engineering applications in Saccharomyces cerevisiae.” FEMS Yeast Res. 12(2) (2012): 197... -
Easi-CRISPR: Generating Knock-In and Conditional Mouse Models
TypeBlog PostPublishedApril 5, 2018, 12:42 p.m....Cre-dependent conditional alleles, has remained a challenge. Enter Easi-CRISPR: a method that harnesses the...First, you’d construct a plasmid with your desired changes, long homology arms and a selection marker, then...template. Although effective, ivTRT may be more challenging for sequences with complex secondary structure...-purified DNA template. Many thanks to Channabasavaiah B. Gurumurthy, Masato Ohtsuka, and Rolen M....PMC5434640 Miura, Hiromi, Rolen M. Quadros, Channabasavaiah B Gurumurthy, and Masato Ohtsuka. “Easi-CRISPR... -
Editor's Choice, October 2016
TypeBlog PostPublishedNov. 4, 2016, 2:30 p.m....had record readership with over 75,000 views. Changing Labor Laws Bring Increased Postdoc Wages Our most...information for anyone considering a postdoc. In “Changing Labor Laws Bring Increased Postdoc Wages,” Future...for some of these institutions to manage these changes quickly and harmlessly. Learn much more about the...Reasons to Use Reddit for Science Communication Changing Labor Laws Bring Increased Postdoc Wages I hope... -
Providing Plasmids to Researchers in Developing Countries: Addgene and Seeding Labs Team Up
TypeBlog PostPublishedApril 5, 2016, 2:30 p.m....to face global challenges, we hope to empower those that need to face these challenges locally. We look...themselves on the front lines of emerging global challenges. However, these same researchers face significant...international distribution As you can see from the chart below, we are making great progress toward spreading... -
The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing
TypeBlog PostPublishedMarch 10, 2015, 2:25 p.m....Antibodies are composed of two light chains and two heavy chains. The ends of the branches contain the...variable, allowing exquisite specificity. This characteristic allows monoclonal antibodies to be used as ...the DSHB, play an important role in scientific exchange. In addition to ensuring affordable access to ... -
RNA Interference in Plant Biology: New Tools for an Old Favorite
TypeBlog PostPublishedOct. 27, 2020, 1:15 p.m....the RNAi trigger delivering the RNAi trigger characterizing the RNAi phenotype. Although all these steps...plants to have enough tissue for downstream characterization. To simultaneously address both limitations...the RNAi output and trivializes downstream characterization, as the entire culture can be harvested instead...Stainier DYR (2020) Transcriptional adaptation: a mechanism underlying genetic robustness. Development 147....186452 Wilson RC, Doudna JA (2013) Molecular Mechanisms of RNA Interference. Annu Rev Biophys 42:217–... -
Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)
TypeBlog PostPublishedJuly 26, 2018, 12:46 p.m....kinase activity is FRET, but FRET reporters are challenging to design and difficult to multiplex. The Covert...which fluorophores can be multiplexed together. Challenging to Design: FRET reporter activity depends on...and NES sequences, which have previously been characterized. See Box 1 in Kudo et al. for more details on...Resources on the Addgene Blog Learn about photoswitchable kinases Learn more about fluorescent biosensors...