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...generating the standard curve for qPCR can be challenging to get right. Here at Addgene, we use a linearized...droplet generation oil are moved through small channels to create a water-in-oil emulsion containing approximately...factor of virus Most droplet readers have a few channels for detecting fluorescence so that it is possible... you should only need to use one. However, two channels can be used to examine the integrity of the viral.../s is recommended to ensure an even temperature change among all of the droplets, but you can go as low...droplet digital PCR for vaccine evaluation in challenged monkeys and TB diagnosis." Emerging microbes ...on the Addgene blog Read more about polymerase chain reaction Browse our blog posts related to viral ...

... Biology (Kimball et al., 2020). Benefits and Challenges CUREs inspire students to take ownership of their...research projects have also been proposed as a mechanism for increasing diversity and inclusion in STEM...experience on their resumes). CUREs are not without challenges. Real research is messy, and it can be difficult...instructors, and provides opportunities for students to exchange ideas and results with peers and faculty at other...win a Nobel!   Jason Pellettieri is Professor and Chair of the Biology Department at Keene State College...(4):602-606. Brian, D. 1996, Einstein: A Life. Chapter 17, Einstein Discovers America. New York./  Callahan...Part I – guided inquiry – purification and characterization of a fusion protein: Histidine tag, malate...

...concept for further development. Figure 2: Mechanisms of deaminase-based adenine transversion editors...editor development going forward. Figure 3: Mechanism of gGBE. Created with BioRender.com. Base ...Acronym Full name Notes   Base transition Base change between purines (A ↔ G) or between pyrimidines ...pyrimidines (C ↔ T).   Base transversion Base change from purine (A or G) to pyrimidine (C or T) or vice-versa...doi.org/10.1038/s41587-023-01821-9 He, Y., Zhou, X., Chang, C., Chen, G., Liu, W., Li, G., Fan, X., Sun, M....M., Miao, C., Huang, Q., Ma, Y., Yuan, F., & Chang, X. (2024). Protein language models-assisted optimization...Li, J., Ren, J., Zhang, X., Zhang, Y., Tang, H., Chang, X., Yu, Y., & Wei, W. (2024). Programmable DNA ...

...base precisely? Until 2016, precise single base changes were only possible through exploiting the homology-directed...efficiently than before. A base editor precisely changes a single base with an efficiency typically ranging...ranging from 25-75%, while the success of precise change via HDR limited to 0-5%. This blog post covers ...wonder if a CRISPR-base editor can perform this change. However, there are no naturally known 'DNA adenosine...used to detect success. Although there is a slim chance of generating indels with base editors, keep in... 627-630. PubMed PMID: 29476916. 10. Kang, Beum-Chang, et al. "Precision genome engineering through adenine...plants (2018): 1. PubMed PMID: 29867128. 11. Li, Chao, et al. "Expanded base editing in rice and wheat...

...light. Activation of these neurons via the channel Channelrhodopsin 2 (ChR2) reduced glutamate transmission... but now we’ll focus on how viral vectors have changed the scope and power of three commonly used behavioral... confirms the previously suggested inhibitory mechanism of locomotor sensitization and might open new ...the open (risky). Taking advantage of these characteristic traits, rodents can be used to study anxiety...mouse to deliver Cre-dependent expression of  Channelrhodopsin 2 (ChR2), a modified receptor protein that...Central PMCID: PMC6213283. Bahi, Amine, Vijay Chandrasekar, and Jean-Luc Dreyer. "Selective lentiviral-...

...cell-type and temporal precision. In ICSS, a test chamber is used in which a nose-poke at a particular hole...but is considerably simpler than ICSS. The test chamber is divided into two halves that are distinguished...animal only receives stimulation in one of the two chambers (see image below). If a circuit is reinforcing...animal will prefer to spend most of its time in the chamber in which it receives the stimulation (conversely...it will spend most of its times in the opposite chamber) [2]. Laser activation is usually controlled automatically...specific behavior but that they more precisely characterized with spatial (CA1), temporal (real-time inhibition... the field benefits from collaboration and the exchange of ideas. The rapid spread of optogenetics throughout...

... problem, how can one little change make a difference? Individual changes to our behavior can lead to ...By initiating little changes in ourselves, we can both influence others to change their behavior and we... Recycling and Innovations posts. Making small changes in your daily habits can feel ineffective or unimportant...Green Lab also has resources and even a freezer challenge that your university could join. Find alternatives...

...this rapidly changing environment. In particular, we are working to understand what challenges remain in ...Addgene saw big changes in 2022, from the launch of a new material to the redesign of a community resource...around the world, are learning to navigate the changes brought about by the pandemic. As we adjust, we...learn from, and a compelling future to plan in the changing landscape of science. We are therefore embarking...to serve the scientific community as we grow and change to meet its ever-evolving needs. For now, we have...

...domain is interesting because by changing certain amino acids you can change the specificity,” explained Zhanar...she hopes will change the intracellular localization of RNA. “You can think of changing bases to other... Abil of the University of Illinois at Urbana-Champaign. The PUF proteins include eight modular repeats...binds a consecutive base in its RNA target. By changing amino acids, it’s possible to produce a protein...

... #1, any change in the normalized intensity of your POI could reflect either a genuine change in your ...your POI, but simply whether its concentration changes in response to experimental parameters. This is... intensity. Researchers use later steps in this chain (blue) to infer or calculate things about earlier... even hit the gel. But what if your experiment changes the expression of your loading control without ...your POI or an opposite change in your loading control. Assumption #2: The loading control can be accurately...become extremely susceptible to error, since tiny changes in intensity can reflect massive differences in...

...see below). Importantly, although their role has changed, restriction enzymes are certainly not dead yet...Introduced in 1977, Sanger sequencing, also known as chain-terminating or dideoxy sequencing, was a major breakthrough...biology98.3 (1975): 503-517. PubMed PMID: 1195397. 2. Chang, Judy C., and Yuet Wai Kan. "A sensitive new prenatal...hybridization for detection of gene copy number changes." Protocols for Nucleic Acid Analysis by Nonradioactive... PubMed Central PMCID: PMC2743315. 5. Roberts, Richard J. "How restriction enzymes became the workhorses...Nicklen, and Alan R. Coulson. "DNA sequencing with chain-terminating inhibitors." Proceedings of the National...: PMC431765. 8. Heather, James M., and Benjamin Chain. "The sequence of sequencers: The history of sequencing...

...Precision genome editing faces the challenges above and more. One chief challenge is upregulating HDR in mature...complex oligonucleotide treatment comes with many challenges, researchers have begun to explore genome editing...approaches for exon skipping. Addgene depositor Charles Gersbach used paired zinc finger nucleases to remove...highest-ranking predicted off-target sites. Each paper characterizes additional, unique facets of CRISPR DMD therapy... the time to clinical approval. Even with the challenges precision editing faces, it’s encouraging to ...Mireault AA, McAnally JR, Li H, Sanchez-Ortiz E, Bhattacharyya S, Shelton JM, Bassel-Duby R, Olson EN. Postnatal...

...Content Accessibility Guidelines (WCAG). These changes mean our content is both available and accessible...expanded a growing collection of thoroughly characterized antibodies, and our contributions to key research...building a portal to easily share sequences and characterization data for over 150,000 plasmids, empowering...closer to making an impact on humanity's greatest challenges. Thank you for being part of this incredible ... (2024). A consensus platform for antibody characterization. Nature Protocols. https://doi.org/10.1038...Cable, H., & Longworth, S. (2024). Antibody characterization is critical to enhance reproducibility in ...

...engineering, Dr. Jennifer Doudna and Dr. Emmanuelle Charpentier. I was joined by Addgene Scientist Matt Ferenc...and Joan Steitz. This award ceremony honored Charpentier and Doudna, who talked about their research leading... leading up to the discovery of the mechanism of dual-RNA-guided DNA cleavage in adaptive bacterial immunity...simpler 1 protein - 1 RNA system Drs. Dounda and Charpentier met at a 2011 conference and decided to collaborate... project started with their labs studying the mechanisms of RNAi, the field for which Craig Mello and ...was a simpler 1 protein - 1 RNA system.  Dr. Charpentier advocated for the crucial role of basic research...

...gel causes the negatively charged proteins to migrate toward the positive charge at the bottom of the gel...and the protein is separated through a matrix by charge. In a denaturing western, the protein is denatured...buffer. These systems, however, require you to purchase expensive system-specific sandwiches.   ...while a western blot detects denatured polypeptide chains. The protein regions available for antibody binding...protein in its native state versus the polypeptide chain. Consequently, antibodies validated for IP might...addition to protein abundance, westerns detect changes to a protein’s post-translational modifications...

... approaches a host cell surface are complex polysaccharide structures, many of which are easily varied... to be explored and exploited. With better characterization, these proteins could function as surrogate...surrogate antibodies for identifying and characterizing sugars and sugar-containing molecules in molecular biology...and pressure. Nebulization units, which can be purchased cheaply, are small plastic containers that can... can perform whole genome amplification (by a mechanism known as multiple displacement amplification) ...cloning site of the vector. In order to reduce the chances of some clones being outcompeted by others in liquid...colonies of interest, sequence the inserted DNA and characterize your new putative host cell-binding proteins...

...researchers involved. Communication is a major challenge when researchers are communicating across disciplines...disciplines or doing different tasks. But these challenges can be exaggerated when communicating across ... help other teams successfully overcome these challenges, too. Schedule regular meetings Scheduling a...person Presenting the results from my lab exchange project at the International Cancer Immunotherapy...other in person. One possibility is to do a lab exchange, where researchers spent time in each other’s ...performed the same way at different sites. I did a lab exchange as part of the collaboration I led. This gave ...

...just how much one’s environment changes them. Addgene gave me the chance to become a real life scientist...wanted to major in biochemistry. There, I had exchanged contact with Amy C., Addgene’s intern coordinator... — such as using Google workspace — and got the chance to work on my very own research project! Coming...scientist. Addgene gave me the chance to grow into a community that had opened its arms to me from day one. .... Addgene has changed my life for the better, and I wouldn’t want it any other way. Toj Toj-Villagran ...

...variable domains of heavy chain only antibodies) (B) Nanomice express hybrid heavy-chain only antibodies that...’s spotlight after researchers discovered heavy-chain only antibodies in the serum of a camel and soon...)!      Figure 1: Comparison of a heavy-chain only antibody and single-domain antibody, under ...need for a species that naturally produces heavy-chain only antibodies. (A) Overview of genetic modifications... way since the serendipitous discovery of heavy-chain only antibodies in a camel. And I suspect we will...

...trees and comparing them with the gene expression changes of more than 100,000 cells undergoing iEP conversion...tags often had similar gene expression pattern changes and reprogramming outcomes, suggesting that outcomes...results suggest that iEP reprogramming is not stochastic, but rather there’s a particular cell state that...successful iEP reprogramming. MettI7a1 encodes an uncharacterized putative methyltransferase. Adding Mettl7a1... expression and clonal history associated with changes in cell identity. The Morris Lab used CellTagging... processes where cells are undergoing identity changes, such as the differentiation of induced pluripotent...