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TypeBlog PostPublishedOct. 25, 2018, 1:23 p.m....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...
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An Introduction to Adenovirus
TypeBlog PostPublishedOct. 3, 2023, 1:15 p.m....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... -
Plasmids 101: Secondary Nanobody Toolbox
TypeBlog PostPublishedFeb. 27, 2018, 2:04 p.m...., Alexa Fluor 568, or Alexa Fluor 647 before application to the sample. Staining with this multi-color...Mouse IgG2a TP1129 pTP1005 (3x cysteines) Mouse Kappa TP1170 pTP1174 (1x cysteine) pTP1172 (3x cysteines...labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation. eLife. PubMed... -
Plasmids 101: Colony PCR
TypeBlog PostPublishedMay 12, 2016, 2:30 p.m....primers. Find Empty Backbones at Addgene Each approach has advantages and drawbacks, which are summarized...which method you choose, make sure to use the appropriate antibiotic for selection. Lysing bacteria...to determine their size. Make sure to use an appropriate molecular weight standard for reference and to... -
CRISPR Antimicrobials
TypeBlog PostPublishedMay 3, 2016, 2:30 p.m....virulent or antibiotic resistant bacteria, and inappropriate antibiotic usage selects for resistant bacteria...to combat antibiotic resistance using multiple approaches. Major restaurant chains including Chipotle and...bacterial host range. To further potential clinical applications, Ando et al. recently engineered synthetic phages... -
Tips for Screening with Yeast Two Hybrid Systems
TypeBlog PostPublishedOct. 22, 2015, 2:30 p.m....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, ... -
Cpf1: A New Tool for CRISPR Genome Editing
TypeBlog PostPublishedOct. 14, 2015, 2:30 p.m...., Zetsche et al. developed a Cpf1 multiplexing approach using a single crRNA array to express up to 4 ...-target editing in a cell culture model. The application of Cpf1 to genome editing is exciting both in...in terms of basic science and translational applications. This discovery of this type V CRISPR system ... -
The Importance of a Fun Workplace: Company Culture at Addgene
TypeBlog PostPublishedSept. 3, 2015, 2:30 p.m.... result seems to be a balance of productive and happy employees. The teambuilding culture helps us build...throughout the years. This team building merriment happened naturally when Addgene was a family affair back... sure people know that there is something fun happening! Addgene actually has chimes outside of our break... -
Hot Plasmids Spring 2024
TypeBlog PostPublishedMay 23, 2024, 1:15 p.m....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... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog PostPublishedJan. 24, 2023, 2:15 p.m.... 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 ... -
Deciphering the Mysteries of Behavior with Viral Vectors
TypeBlog PostPublishedSept. 17, 2019, 12:40 p.m....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... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog PostPublishedNov. 19, 2020, 2:15 p.m....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... -
Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)
TypeBlog PostPublishedJuly 8, 2025, 1:15 p.m.... 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 PostPublishedMay 20, 2025, 1:15 p.m....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... -
AAV Titers: Where Do They Come From And What Do They Mean?
TypeBlog PostPublishedNov. 15, 2017, 3 p.m....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 ... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog PostPublishedOct. 19, 2017, 1:54 p.m.... 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... -
Changing Labor Laws Bring Increased Postdoc Wages
TypeBlog PostPublishedOct. 25, 2016, 2:30 p.m....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 PostPublishedMay 5, 2016, 2:30 p.m....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 Optogenetic Tools for Cytoskeleton and Membrane Control
TypeBlog PostPublishedSept. 28, 2023, 1:15 p.m.... 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... -
Harnessing Bacterial Toxins for Allelic Exchange
TypeBlog PostPublishedAug. 15, 2019, 12:30 p.m....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...