... #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...

...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...

... 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...

... material, and variations between serotypes can change how the viral particle sneaks its way into cells...Citation CNS AAV1, AAV2, AAV5, AAV8, AAV9, AAVrg Aschauer et al., 2013 Heart AAV9 Bish et al., 2008 ...expertise! References and resources References Aschauer, D. F., Kreuz, S., & Rumpel, S. (2013). Analysis...., Yin, L., Merigan, W. H., Flannery, J. G., & Schaffer, D. V. (2013). In vivo-directed evolution of a...Y., Jae, L. T., Wosen, J. E., Nagamine, C. M., Chapman, M. S., & Carette, J. E. (2016). An essential receptor...Gaj, T., Lavzin, M., Ritola, K. D., Lindo, S., Michael, S., Kuleshova, E., Ojala, D., Huang, C.-C., Gerfen... Dudman, J. T., Hantman, A. W., Looger, L. L., Schaffer, D. V., & Karpova, A. Y. (2016). A Designer AAV...

...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...

...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...

...Leiwe and Takeshi Imai from Kyushu University. Stochastic multicolor labeling is a powerful solution for...microscopy-based neuronal reconstruction. To achieve stochastic multicolor labeling, Brainbow used the Cre-loxP...copies of the transgene cassette are introduced, stochasticity will result in a combinatorial expression of...Brainbow was inherently low. This is because the stochastic and combinatorial expression of fluorescent proteins... such vector-based gene delivery systems, the stochastic expression of XFPs is possible by simply introducing...introducing a mixture of XFP genes. Stochastic multicolor labeling is possible when each of the XFP genes...

...However, these approaches are often met with the challenges of regulatory bottlenecks and their scope is ...is limited by the lack of well characterized inducible promoters. Far removed from the bio-industry’s vision... various metabolites in Escherichia coli or Saccharomyces cerevisiae with few techniques that are easily...designed scaffold binding sites. With hundreds of characterized aptamer domains available, an unprecedented .... This strategy can be applied to effectively change metabolic flux. The dCas9 protein and orthogonal...6441/ Resources: An open source library of characterized aptamers and supporting publication: http://...

...they change color as they shift from "off" to "on" in the presence of calcium. This color change can be...problem that he considers “one of the greatest challenges of neuroscience.”  Hot plasmids: Twitch sensors...imaging neurons, Griesbeck adds that this color change has a particular advantage for observing events...sensitivity. As reported in Nature Methods, the FRET changes of these Twitch sensor variants were optimized ...trial and error. Transition metals also present challenges in that they are found in cells at much lower...

...through filtration methods because of its ability to change shape and the absence of a rigid cell wall. Here.... For testing, the medium should not have been changed for at least two or three days before sampling ...chemical reactions and this induces a noticeable color change. This method has high sensitivity but low specificity...specific enzymes are used which trigger cellular mechanisms that generate luminescence. This can then be ...are working without gloves or masks. Make sure to change the gloves and masks regularly instead of keeping...water provided to maintain the humidity should be changed regularly as mycoplasma, bacteria, and fungus can...Mycoplasma Contamination in Cell Cultures by Polymerase Chain Reaction. In: Methods in Molecular Biology. Humana...

...questions in the chat box. That’s another benefit to note-taking: you can revisit the Q&A in the chat box. Presenting... plan more with regard to my interests.” Use the chat box and upvote If the thought of walking up to the...asking questions using the polling feature or the chat box. One key thing to remember regardless of the...poster,” he says.Viewing a poster on a screen is challenging, requiring you to zoom in to different parts ...than in-person. You might miss out on the short chats between sessions or waiting for a session to start...

...years. They helped one another navigate career changes and overcome barriers by sharing broader perspectives...Healthcare Businesswomen’s Association (HBA) Boston Chapter and the Massachusetts Association for Women in ...diversity of input and perspectives Provides a good mechanism for accountability that makes mentoring effective... heard from peer groups meeting at both the Fox Chase Cancer Center and Brandeis University. 5 easy steps...choosing a topic and leading the discussion. This is a chance to practice leadership and communication skills...better with process, so don’t just get together and chat. Have an agenda and get everyone ready for the discussion...

... simultaneously   Researchers in the labs of Michael Lin and Ryohei Yasuda have teamed up to develop...maturation. Three new RFPs were isolated and characterized: mScarlet (fluorescent lifetime of 3.9 ns, quantum...member of this class that works via a different mechanism - the PhoCl protein is irreversibly cleaved when... also provided a detailed protocol at Protocol Exchange. Plasmid maps can be found at Addgene and in Figure...Gram-positive bacterium, many of its genes are poorly characterized. The Gross lab hopes these libraries will be...Transcription factor fusions elucidate iPSC reprogramming mechanism The Ptashne lab was the first to demonstrate ...used this now classic technique to elucidate the mechanism by which SOX2 acts to reprogram human fibroblasts...

...Creating epitope-tagged clonal cell lines remains a challenge as it relies on inefficient homology-directed ...representing a protein super family). To overcome these challenges, we set out to develop a hassle-free, highly ...Technologies, USA). These synthetic RNAs can also be purchased from various other suppliers with a typical turnaround...straightforward as the RNA molecules can be directly purchased and there is no need to construct plasmid vectors...; this was possible because reagents could be purchased directly from commercial suppliers without investing...template) and use homology arms of 60 -70 bp . Purchase gene-specific crRNAs and matched ssODNs from commercial...by single-cell deposition in 96-well plates: Characterize clonal lines by ICC and PCR genotyping (Sanger...