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...thus tracking the localization of that biomolecule inside living cells. FPs classified by the emission color...range, there are other traits that need to be considered when choosing an FP: Unique categories of fluorescent...distance between the two FPs is <10nm, and should be considered when labeling proteins that interact. Indeed,... coefficient and quantum yield of the protein, divided by 1000. In many cases the brightness is compared...greenfluorescentblog.wordpress.com.   More Helpful Websites & Resources: FP guide at Addgene Interactive Visualization of Fluorescent...

... degree of identity, so not suited to generate polycistronic containing multiple identical polyadenylation... can be assembled. For example, because CpG dinucleotides are underrepresented in most vertebrate genomes...and MluI recognition sites, which contain CpG dinucleotides in their 6-mer sequences, are rarely found in...for cloning genes from organisms with high CpG dinucleotide representation. The specific restriction enzymes...

...Heinrich, W. (1983). Worldwide epidemiology of human adenovirus infections. Am J Epidemiol, 117(4), 455–466...length. Human adenovirus type 5 (hAdV-C5) can be considered the prototype, although many viruses infecting...adenoviruses zoonotic? A systematic review of the evidence. Emerg Microbes Infect, 8(1), 1679–1687. https...

... but many find it helpful to obtain support and ideas from those who seem to be managing. Being reminded...balance tips, we need to look to for support and ideas to manage our stress. If you are saying to yourself...appreciate this – and they will be happy too. On the flip side, if you are not happy at work, you probably need... you will get help when you need it. On the work side, join a professional community that you can count...an emergency and this will make you much more confident about their safety when they leave home which ...

...ternary complex with the multiple myeloma drug mezigdomide and ubiquitination substrate Ikaros (IKZF1ZF2...important functional properties, and is amenable to a wide range of biophysical assays (Kroupova et al., 2024...CRBNmidi as a promising tool for future structure-guided drug design and studies of protein ubiquitination...showed greater photostability and also matured considerably faster, with expression levels comparable to...expression and removing the sgRNA plasmid after identifying successful knockouts limits further cutting and...

...negative staining. Empty vector particles can be identified after negative staining and appear darker than...been used to titrate AAV, but this method is not widely used today (Fagone et al., 2012). PCR-based methods...from different sources in the same experiment, consider retitering both AAV in your lab. The absolute ...does not give an indication of infectious titer, consider titrating each lot of AAV in vivo to determine...Additional Resources on the Addgene Blog Important Considerations When Using AAVs Addgene's AAV Quality Control...

... you have these rules (word limit, formatting guidelines, etc.) you have some criteria to shape the document...research papers that I needed to read. Once I had an idea of the themes in the field, I searched for recent...field. I made sure to find information from genome-wide studies, as well as results from smaller and more... or popular papers, but looked for papers from a wide range of authors. Just start writing When I first...Plasmids Review the CRISPR Field with Our CRISPR Guide Pages ...

...Sequencing results are analyzed to confirm the identity and integrity of the viral genome and the absence...Samples of viral preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver...negative staining. Empty vector particles can be identified after negative staining and appear darker than... monitored throughout production, ensuring the identity of each sample from the DNA tube all the way to... preparation. After production, we confirm the identity of the transfer plasmid in the viral preparation...

...Mortensen et al., 2019). Our method might provide some useful guidelines for the development of transient transformation...transformation systems for other species as well. Things to consider in developing a transient leaf transformation ... promoter in Agrobacterium. This can be easily avoided by the use of introns, which will be removed from... to quickly assemble vectors and test different ideas. We are a huge fan of the Golden Gate based MoClo...

...generating interesting drug-like compounds especially cardenolides which have been used to treat heart disease....disease. Caterpillars that eat milkweed generate cardenolides that tastes yucky (and is poisonous) for bird ...that evolved in the last 10,000 years (which is considered recent). They have diverse traits with good genetic...control who a female wants to mate with and if she decides to raise their offspring or leave them to their...a cool fluorescent indicator system to rapidly identify recombinants. Arthropods (Jason Rasgon) This lab...

...Hedgehog but this is not a direct reference to the video game character but rather to an extremely important...Christine Nusslein-Volhard and Eric Wiechaus first identified the hedgehog gene in Drosophila. They observed...nonsyndromic holoprosencephaly where the brain fails to divide properly into the right and left hemisphere. Addgene...origins of the name but some of our Addgenies had an idea: sqh mutant fly ovaries had “squiggly” actin fibers...Additional resources on the Addgene blog Read our Quick Guide to Working with Drosophila Read our Plasmids 101...

... one bacterium sharing genes with another, an accidental transmission of genes from one bacterium to another...tried and true process of evolution. Watch this video showing bacteria evolve antibiotic resistance on...different reasons. Exposure to antibacterial drugs provides a selection pressure, killing off bacteria that...prescribed to them This may seem counter-intuitive considering the other three scenarios, but a short treatment... antibiotics and which ones aren't?     This video shows how to perform a Kirby-Bauer disk diffusion...

...post on viral vector parts. Promoter specificity Aside from choosing a promoter based on type of RNA transcript...transcript     Description Expression Additional considerations CMV General expression mRNA Strong mammalian... phosphoglycerate kinase gene.  Constitutive  Widespread expression, but may vary by cell type. Tends ... used for Description Expression Additional considerations T7 in vitro transcription/ general expression...therapeutic uses, and in those cases it's important to identify the right tissue-specific promoters as described...

...Addgene's Viral Vectors Guide Addgene's Chemogenetics Guide Addgene's Optogenetics Guide  ... our Viral Vectors 101 eBook!  This practical guide to using viral vectors in the lab is a helpful resource...

...promoter Mature siRNAs typically range from 19–22 nucleotides and are 100% complementary to their target sequence...plasmid and call it a day. You need to include both sides of the complementary RNA duplex and the hairpin ...169)","rgba":"rgba(0, 82, 169, 0.75)"},"override_default_style":true,"text_hover_color":{"color":"#ffffff...time you need to knock down a gene, maybe you’ll consider using RNAi! References and Resources References... R. (2021). Reexamining assumptions about miRNA-guided gene silencing. Nucleic Acids Research, 50(2), ...

...so we’ve been busy updating our CRISPR guide. Our CRISPR guide is a comprehensive resource that covers...week at #AddgeneCRISPRweek. We updated the CRISPR guide! If you’re not into Twitter, we have some exciting...for planning your CRISPR experiment. For a quick guide to CRISPR, download our CRISPR Cheat Sheet....

... Summers, 1998). Multimeric plasmids are also considered to be less stable and more prone to being lost...other methods below should help in multimer identification.   Fig. 2: Gel image of Plasmid 61564...used to detect plasmid multimers. Some companies provide NGS whole plasmid sequencing services using long-read...within a plasmid and, therefore, allow for the identification of plasmid multimers. Capillary Gel Electrophoresis...laser-induced fluorescence (CGE-LIF) for detection and can provide data on the quantity of various topologies and ...

...this concept in his microbiology lab course, he decided to add a bit of fun. He mixed together 10 E. coli...host strains affect transformation efficiency.  Aside from the Streak Plate Challenge, Westenberg’s course...plasmids here, and check out our streak plate protocol video on Youtube:     Additional resources on the...plasmids at Addgene Check out additional written and video protocols ...

...insights into the process of getting an MD PhD, the inside scoop on developing new CRISPR tools, and the potential... : Alex introduces himself and explains why he decided to do an MD PhD - it's all in the family. 3:56 ...Biologically Inspired Engineering and why Alex decided to work there. 13:22 - 22:07 : Alex's projects ... Resources on Addgene.org Check Out Our CRISPR Guide Pages Find Ready-to-Use CRISPR Lentivirus Learn ...

... In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However...compatible with the introduction of unique molecular identifiers, which can improve the statistical power of in... of the pRDA_526 vector. Moreover, the screen identified putative novel regulators of CD8+ T cell responses...Context-Dependent Germline Knockout in Immune Cells Genome-Wide Screening Using CRISPR Cancer and the Immune System...