We narrowed to 858 results for: CAL;

...allows for directional assembly of DNA sequences, called modules or parts. Assembly is dictated by four-...for fresh bacteria. Performing your Assemblies Calculate your volumes As you progress through the MoClo...increase your ligation efficiency, we recommend you calculate the right volume of each plasmid according to ...the equation in Figure 3.    Figure 3: Calculation for the plasmid volume for MoClo experiments....high-throughput, we recommend transforming into chemically competent cells in a 96-well PCR plate. Always...

...production of viral vectors typically involves shuttle vectors, also sometimes called transfer vectors. The ...antibiotic works. Another example is Zeocin, a DNA intercalator, which affects all kingdoms of life. Of course...host range plasmids shuttle vectors? While it’s technically accurate to say that shuttle vectors have a “... “broad host range,” these terms are typically used in different (though overlapping) ways. Broad host...

...product purity of 66%. Pro tip! Abasic sites are also called apurinic or apyrimidinic sites and abbreviated ...are the same protein!) (Chen et al., 2024). They called their initial editor AXBE, for A → X base editor...in a G → Y (Y = T or C) base editor, which they called glycosylase-based guanine base editor (gGBE) (Tong... Therapeutics team has deposited their editors, called gTBE and gCBE, with Addgene. gTBE can introduce...site A DNA site with the nucleobase removed. Also called an apurinic / apyrimidinic (AP) site. UNG / UDG...

...regions and allocate specific functions to them. Electrical recordings and lesioning studies have been essential...within the LC of mice. When those neurons are chemogenetically activated, norepinephrine release causes a...an enhanced propensity for movement and activity called locomotor sensitization. This enhanced movement...responsible for the antidepressant effect. To specifically activate the Drd1 neurons, they injected AAV2... "Hypothalamic CNTF volume transmission shapes cortical noradrenergic excitability upon acute stress."...

...molecules, forming a “handcuff” structure that physically blocks replication machinery when plasmid copy...It is also worth noting that plasmids can be symmetrically incompatible, meaning that both plasmids are... from the cell with the same probability, or asymmetrical, where one plasmid lost from the cell at a higher...deal with the more sinister plasmids of the microbiological world. (Want to know more about these plasmid...high copy incompatible plasmids that lead to asymmetrical plasmid loss of the usually large, low copy ...

...experimental design and its potential for clinical applications. Specifically, efficacy rates of Cas7-11 were reported...current knowledge. The Abudayyeh-Gootenberg lab (also called the AbuGoot lab), a joint lab run by Omar Abudayyeh...originally suspected that this new Cas7-11 might be phylogenetically classified as the more complex Class I systems...switched up the serine integrase for one which can enzymatically take a genetic sequence and integrate it into...

...JAT, Callaway EM (2007) Monosynaptic Restriction of Transsynaptic Tracing from Single, Genetically Targeted...Why pseudotype? Lentiviruses and retroviruses typically target immune cells with their wild-type envelopes...out this review article published in Reviews in Medical Virology (Li et a., 2018).   Figure 2:...6178.1997 Wickersham IR, Finke S, Conzelmann K-K, Callaway EM (2006) Retrograde neuronal tracing with a deletion-mutant...

...studying biochemistry at the University of Southern California. He is also an intern at the Yang Lab of the ...transposon-encoded CRISPR/Cas systems in bacteria, specifically on optimizing strains with low efficiency.   ...of Sciences. He joined Shanghai Institutes for Biological Sciences in 2000 and became a professor in Synthetic...reprogramming of cells to understand the genetic and biochemical mechanisms behind their distinct phenotypes and...generation of engineered cells for green chemistry and biomedical applications.   Yiwen Zhang entered the Sheng...

...expression limiting regulatory elements or use a non-canonical code (Gustafsson et al., 2004), such as Hoogsten...recognize multiple synonymous codons) may actually be critical for determining the rate of translation. The ribosome...10.1101/gr.139758.112 Mauro VP, Chappell SA (2014) A critical analysis of codon optimization in human therapeutics... GluCl ion channel genes for mammalian cells dramatically improves expression levels. Journal of Neuroscience...synthetic? How Addgene;s dataset reveals trends in biological innovation Resources on Addgene.org Check out...

...In Addgene’s early days, we would ‘spot-check’ critical regions of plasmids deposited with us via Sanger...development teams, frequent updates allowed us to scale our lab work as we grew. As the number of deposited...us to switch between executing our pipeline on a local computer or in a cloud environment. Given the throughput...jumping back and forth between running the pipeline locally and in the cloud. Creating a bioinformatics workflow...script file (containing our new process) on our local computer. Eventually, our workflow would handle ...

...post was updated on Jul 27, 2020. CRISPR, and specifically Cas9 from S. pyogenes (SpCas9), is truly an ...specified by the crRNA sequence, it adopts an enzymatically “active” state rather than reverting to an inactive...propensity of Cas13a to bind and cleave RNA non-specifically. The ability of dCas13a to bind target RNA without...modulating splicing, and regulating expression of specifically targeted RNAs. Finally (and perhaps most interesting...observed shRNA off-targets. They subsequently used catalytically inactive dLwaCas13a as a programmable RNA-binding...

...the Janelia Research Campus of the Howard Hughes Medical Institute. Are you currently on or planning to ...rejection of a model and no clear alternatives. Typically, a research vision is broken up into three aims... basis of a single grant. Either way, think realistically about the time and resources required to feasibly... aims in more depth in the remaining time. The typical chalk talk structure is: What is the big question...the Janelia Research Campus of the Howard Hughes Medical Institute. His interests include the quality control...

... this post I’d like to delve deeper and look specifically at the characteristics and capabilities an ELN...facilitate true reproducibility. The following are critical: (1) connectivity to other data sources used in...likely stored in institutional and lab servers specifically designed to hold large amounts of data. Although...group’s research – it makes no sense and is not practical to move all this data in bulk into a lightweight...lightweight tool like an ELN. It is nevertheless critically important to be able to conveniently reference specific...

...FRET during a journal club, my guitarist brain automatically thought about the raised element found on the...interactions in living cells using a microscope. Historically, molecular interactions were detected by indirect...know or interact with each other. FRET reduced the scale of our perception about molecular interactions. ...Periasamy defined FRET as "a distance-dependent physical process by which energy is transferred non-radiatively...FRET) microscopy imaging of live cell protein localizations. J Cell Biol. 2003 Mar 3;160(5):629-33. ...

... RT-qPCR tests to diagnose COVID-19 and some serological tests to determine past exposure. However, the...rapid detection.  Highlights: Cas13a is not catalytically active in the presence of two or more mismatches... copies RNA targets in a 40 min incubation. A scalable, easy-to-deploy, protocol for Cas13-based detection...available and less expensive. Instead of using the typical PCR machines, the authors use affordable, field-ready..., Acosta-Alvear, D., & Wilson, M. Z. (2020). A Scalable, Easy-to-Deploy, Protocol for Cas13-Based Detection...

...set of transferable skills, coupled with your technical skills, can open new career opportunities for ...conflict can also be good. In science, a reviewer’s critical comments on your manuscript is a form of (hopefully...scientist. And some amount of conflict is actually critical in the business world. Companies and organizations...aspects of customer support each team “owns” is critical for day-to-day operations and keeping our customers...especially people with whom you strongly disagree, is a critical component of conflict resolution. Having empathy...

...insights into the complicated physical organization of the brain and other biological systems. Interested in ...deletions are defined using 2 loxP variants (only identical loxP sites can mediate recombination.) The deletions... preventing further recombination. Brainbow-1.1 scales up the system to four fluorescent proteins, with...homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Snippert HJ, van der...

...without their problems, but much genetic and physiological data have been accumulated over the years using...check out our Mouse Modeling blog posts: Part 1: Genetically Engineered Mice and Part 2: Breeding and Crossing...into the oviduct of pregnant females, a technique called Genome editing via Oviductal Nucleic Acids Delivery...small mutations with HDR, when it comes to larger scale genome editing, such as replacement of a mouse gene...nice to your microinjectionist, who has the strategically important job of delivering the payload! Last...

...lab, or tissue culture as it is commonly called, is a critical tool for many scientists. Mammalian cells...AAV preparation. For detailed instructions and calculations on how to prepare a plasmid stock used to generate...cells provide scientists with the means to study biological processes on the cellular level instead of having...

...completing her PhD in Chemistry and Chemical Engineering with Harry Gray at Caltech, Crystal moved on to a Postdoc...Investigator III at Novartis, a multinational pharmaceutical company based in Switzerland. After completing...scientific teams at the Novartis Institutes for BioMedical Research to develop novel biologics for the treatment...

...Addgene podcast, we sit down with Alex Chavez, a Clinical Fellow in Pathology at Masachusetts General Hospital...Church and Collins labs at the Wyss institute for Biologically Inspired Engineering. Alex is the first MD PhD...and explains why he loved the first 2 years of medical school. 7:05 - 10:55 : Alex's focus during graduate...postdoc 12:01 - 13:21 : The Wyss Institute for Biologically Inspired Engineering and why Alex decided to...

...use as a genetic engineering tool is frequently called recombineering - short for homologous recombination-mediated...insert site (Figure 2, top).  These primers are typically ~70 nucleotides long (20 nucleotides that anneal...fragments, such as gene replacements and tags.  The typical frequency of recombinants is 1 positive clone out...efficiency than the other and you’ll find out empirically which is the lagging strand. ssDNA substrate ...below).  Controlling expression of Red proteins is critical for minimizing the toxic effects of Gam expression...

...networks in biological systems. NGS will also play significantly into the realm of clinical therapeutic...validation process, also known as CRISPR genotyping, is critical to demonstrating causal relationships between ... forms of off-target analysis in order to meet clinical approval. It is key to maintain a consistent set...the detection of KRAS mutations: diagnostic and clinical implications." The Journal of Molecular Diagnostics...

... is necessary for bacterial transformation of chemically and electrocompetent cells is minimal (picogram...nucleic acids (genomic DNA and RNA), proteins and chemical contaminants that have been used during the purification... preparations for this technique must be larger scale, to allow the extraction of enough plasmid to last...addition to being free of any nucleic acid and chemical contaminants, these DNA samples must also be entirely...

...in Plasmids 101, tags can help you determine localization of a protein of interest, purify it, or determine...major caveat - a tag may interfere with protein localization and/or function, so each tagged protein must...proteins found that 20% of tagged proteins did not localize similarly to their untagged counterparts. These...fluorescent-protein tagging show high correlation for protein localization in mammalian cells." Nature methods 10.4 (2013...

...metabolites that are dynamically secreted by muscle during exercise to meet the brain’s physiological demands,” ...protects the brain from depression. Mice that are genetically modified to produce extra PGC-1a1 only in their..., the brain is protected from stress-induced biochemical changes that could otherwise lead to depression...

...Emeritus at UCSF. Dr. Bourne has some radical (and some not so radical) suggestions for defining the problem... training too many scientists and some propose radical changes to the way we view the graduate school ...University founded a successful consulting company called The Balsa Group. University of Pennsylvania has...

...for reference: Untergasser Lab Preparation of chemically competent cells (protocol) Preparation of electrocompetent...until we are ready to perform transformations. A typical 200mL bacterial culture produces 20mL of competent...protocols below: Untergasser Lab Transformation of chemically competent cells (protocol)  Transformation of...plasmid is introduced to bacteria, the bacteria is typically grown in liquid culture without antibiotics to...

...paper-based, affordable, and portable microscope called the ‘foldscope’ (Dr. Prakash, Stanford University...as conventional microscopes and give a similar optical quality (magnification of 140X and 2 micron resolution...I have worked many hours in a dark room with a confocal microscope, yet I was once again fascinated by...them. Addgene depositor Dr. Mikhail Shapiro from Caltech presented an exciting way of listening to microbes...

...excellent Technical knowledge and creativity Become a “thought leader,” develop a technical “niche,” develop...FierceBiotech to “learn the business” Attend events with local networking and trade organizations Another excellent... be found on the UK Career Site for Researchers called Vitae. Vitae recently published a career development...

...pride in facilitating the exchange of not just the physical samples, but also the legal documentation that... Addgene has adopted the terms of the Uniform Biological Material Transfer Agreement (UBMTA) for its material... may not use the material in human subjects or clinical trials Acknowledgement in recipient’s future...have opted into our eMTA system can even sign electronically or choose to auto-approve MTAs. Whatever your...

.... All of this requires genes to be regulated dynamically.” To understand how that works in memory formation...they could package LITEs in viral vectors and genetically target them to specific cell populations. They... the genetic and epigenetic players in normal biological processes and disease states. While LITE plasmids...with LITE? Reference: Silvana Konermann et al. "Optical control of mammalian endogenous transcription and...

...Antibody characterization is critical to enhance reproducibility in biomedical research. eLife, 13, e100211...launched AAV Packaged on Request, a service that dramatically expands and simplifies your viral vector options..., K., Alshafie, W., Baker, M. R., Ball, A. R., Callahan, D., Cooper, J. A., Crosby, K., Harvey, K. J.,...

... therapies,” said Kris Wood of Duke University Medical Center. “While we do not yet have a comprehensive...resistance to drugs,” in a rapid, straightforward, and scalable manner, Wood said. Thankfully, the researchers...generate a kit which is now available at Addgene. “We called it cloning by mail,” Wood says, and it took them... he said. “It could [be used to screen for] morphological features, migration, or survival in different...

...while - a tool to expand perception, which is ironically what science is about. For me it is essential...living organism I study, to be able to see it all scales. Most modern biologists focus on one aspect alone... time, defend the thesis, etc. Photography is critically important in many fields of biology and I believe...appreciated by a wider audience for more than just its biological importance. More of Daniel's stunning work can...

... Connie Cepko (Harvard Medical School), and Dr. George Church (Harvard Medical School) for taking the ...the lab with parties in the lab’s common areas, local pubs, and homes. The Church lab also toasts a newly... Improving your work-life balance Balancing the Scale: NSF's Career Life-Balance Initiative Can Scientists...

...notebooks and binder on your desk. Arrange them in chronological order (or by project, if that works better for...upcoming grant deadlines, and sync everything to your calendar. Ditch the paper To Do List and move to one that...reminders. These programs can be synced to your online calendar and can be accessed from your computer or smartphone... notebooks (ELNs) have gained traction with pharmaceutical companies, academic labs have been slower to...

...nearly all domains of life repair genomic damage, specifically double strand breaks. Researchers have long ...by direct transformation. A given insert will typically contain both a protein modification sequence and... not only track the abundance, movement, and localization of individual proteins, but also investigate... green, and red FPs that are codon optimized specifically for expression in yeast. These tagging vectors...

...expression of the indicators over white dashed lines. Scale bars, 10 μm (HEK293T), 20 μm (neurons). Figure and...feedback in response to norepinephrine release, and clinically-used drugs targeting these receptors cannot be...nLightR, both generating a measurable signal after electrical stimulation with a single pulse, while nLightR...Cui, G., Lin, D., Du, J., & Li, Y. (2019). A Genetically Encoded Fluorescent Sensor for Rapid and Specific...

...imaging method that uses iterative staining and chemical bleaching to allow for high-resolution imaging...not recommended reagents) along with information critical for the performance of a reagent, e.g., antigen... (2022). IBEX: An iterative immunolabeling and chemical bleaching method for high-content imaging of diverse...Germain, R. N. (2020). IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial...