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TypeBlog Post...which a vaccine is available. However, here it appears that the culprit for these unusual cases was either... the removal of certain adenoviral genes that apparently contribute to virulence (Tollefson et al., 1996...on the Addgene blog Viral Vectors 101: Viral Applications Adenoviral Vector Production and Troubleshooting...
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Tips for arabidopsis transformation
TypeBlog Post...Addgene Choosing an expression system Choosing an appropriate expression vector is the first step in generating...binary gateway vectors and can be used for promoter swapping and generating N and C terminally tagged fusion...phenotypes. For most phenotypic analysis it is appropriate to begin working in the T2 generation. The selection... -
Tips for Screening with Yeast Two Hybrid Systems
TypeBlog Post...or growth stage). A prey library can either be applied to the bait in a single population (e.g. transforming...although these can often be accounted for using appropriate replicates and controls (see Tips below), and.... Obrdlik, P., El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, C., Ellerbrok, ... -
AAV Titers: Where Do They Come From And What Do They Mean?
TypeBlog Post...used to generate AAV. One study showed that approximately 50% of viral capsids were genome-containing ...particles can be identified after negative staining and appear darker than full vector particles. This image shows...experiments? Because (physical) AAV titers are approximations and they depend highly on the experimental ... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog Post...highlights some of the latest breakthrough therapeutic approaches. What is Rett Syndrome? Caused by a mutation ...technical challenges to overcome for both of these approaches, such as delivery of the editing machinery to...Extends the Survival of MeCP2-Null Mice without Apparent Toxicity after Intracisternal Delivery. Molecular... -
Deciphering the Mysteries of Behavior with Viral Vectors
TypeBlog Post...movement in an open field box or a circular corridor apparatus using a camera or tracking software. Locomotion...and might open new ways for a novel therapeutic approach against drug addiction. Anxiety tests Rodents...measuring the time spent in a particular part of the apparatus, it turned out that the IL-1 beta overexpressing... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog Post... its reintroduction, inversion and re-inversion happen at the same probability. Continuous inversion of...the same time from two different plasmids this approach becomes problematic as it reduces the probability... Cell Types for Imaging and Long-Range Circuit Mapping.” Journal of Neuroscience 28(28): 7025–30. PubMed... -
Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)
TypeBlog Post... process, and request your sequence with the appropriate flanking restriction sites and overhangs already... help you in your MoClo quest, so good luck and happy cloning! Resources Additional resources on the...Modular Cloning Plasmids 101: Modular Cloning Applications and Kits Plasmids 101: Golden Gate Cloning ... -
Plasmids 101: Shuttle Vectors
TypeBlog Post...which helps increase cellular uptake, but this approach requires compatible cell lines that express viral...individual analysis. A shuttle vector designed for this approach in a mammalian cell experiment should still include...terms are typically used in different (though overlapping) ways. Broad host range plasmids are those that... -
Changing Labor Laws Bring Increased Postdoc Wages
TypeBlog Post...institutions, or institutions they're thinking about applying to, are doing - for example, your benefits may...get rid of postdocs; that current postdoctoral appointments may shorten in length; that postdoc salaries...trainee workforce has clearly not been a sustainable approach to biomedical research in the U.S. and postdoctoral... -
The Challenges of Cell Culture
TypeBlog Post...especially with industry. Today we also have the appropriate tools to verify our cells lines. Even if you ...the results are reliable. With this two-pronged approach, the many mistakes of the past can be prevented...including GSK, AstraZeneca-MedImmune, and Infors & Applikon. Due to his high level of experience he now travels... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog Post... the bacteria were injected, limiting their applicability. They also did not scatter ultrasound nonlinearly...chromatic-opening agents to boost expression to appreciable levels. We wanted to create second-generation...ultrasound imaging of nanoscale acoustic biomolecules. Appl. Phys. Lett. 110, 073704 (2017). Hurt, R. C. et ... -
Hot Plasmids Spring 2024
TypeBlog Post...Paul Blainey’s lab has developed a multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi...for microglia research and broader therapeutic applications. Find microglia-targeting AAV plasmids here...reconstituting coding sequences for a range of applications, including CRISPR activation by dCas9-VPR, prime... -
Harnessing Bacterial Toxins for Allelic Exchange
TypeBlog Post...References Espah Borujeni, Amin, Anirudh S. Channarasappa, and Howard M. Salis. "Translation rate is controlled...Scarless Modification of Proteobacterial Genomes." Applied and environmental microbiology (2019): AEM-00990...chromosomal gene replacement in Escherichia coli." Appl. Environ. Microbiol. 81.15 (2015): 5103-5114. PubMed... -
Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection
TypeBlog Post...study its function is one of the most popular applications of fluorescent proteins. These fusion proteins...targeted to the endoplasmid reticulum based on the appearance of OSER whorls. This assay is quantitative and...linker sequence short In the early days of GFP applications, many were concerned with steric hindrance by... -
The Fluorescent Vegetables in Aptamer Soup
TypeBlog Post...enjoy a warm meal on a winter day, you should be happy to learn that many DNA-based reagents are named ...aptamer soup by exploring the development and applications of fluorescent RNA aptamers in Addgene’s collection...fluorescence when bound to DFHBI despite being approximately half the length (Warner et al., 2014); its length... -
CRISPR 101: Validating Your Genome Edit
TypeBlog Post... explain how to verify that your cells were appropriately edited for your desired mutation - insertion...frequency is 50%, then you could infer that approximately 25% of cells are likely -/-, 50% are -/+, and...sequencing confirmation. All knock-ins A method applicable to knock-ins both large and small is a software... -
Multiple Plasmids at a Low Price: Kits from Addgene
TypeBlog Post...polycistronic E. coli expression constructs. Some applications of plasmids from this kit include the production...can be engineered to bind specific RNA bases. By swapping out these different modular repeat segments, you...variety of functions via different fusions. Some applications of PUF protein fusions reported in the literature... -
New Optogenetic Tools for Cytoskeleton and Membrane Control
TypeBlog Post... barrier of the endothelium, but they could be applied to study other actin-dependent processes in cells...) and to introduce unnatural head groups for applications like fluorescent labeling via click chemistry...constructs are great for in vitro biocatalytic applications where toxicity doesn’t matter. Plus, for folks... -
Technologies Enabled by NanoLuc® Luciferase
TypeBlog Post...et al (2). In this post, I’ll cover two great applications of the NLuc technology. The first couples NLuc...the coupled luciferase. This strategy was first applied using RLuc8, an engineered Renilla luciferase, ...fluorescence. The paper reports a wide variety of applications of the Enhanced Nano-Lanterns including monitoring...