...the region of the brain where DA quantitation is desired (Figure 1). The probe perfuses liquid, which causes...et al., 2018). These sensors also exhibit high magnitudes of fluorescent responses. Studies on the kinetics...Ultrafast neuronal imaging of dopamine dynamics with designed genetically encoded sensors." Science 360.6396...

...ve just discovered that your favorite gene was described in a CRISPR screen publication. You see this mentioned...This format is inherently flexible and allows the broadest possible scope of screens to be curated, for example...other than cell viability. The following video provides an in-depth walk through of how to use ORCS:   ...

...instrument. They are the oldest sibling in a Guatemalan family of Mayan descent.   ...teaching me the basics of physics and sustainable design while allowing me to do my internship in the daytime...

...in the initiation of a vast number of signaling cascades, inducing different cellular responses. One of...localization of the Rluc8 insertion within the Gα. Despite exhaustive refinement, it was not possible to generate...protein signal transducers. The TRUPATH collection provides scientists with a toolkit to gain an understanding...

.... MBP has a similar function as GST in that it provides a method to express a foreign protein in bacteria...protease. The carboxyl-terminus of the fusion protein includes an AviTag and a 6X HisTag for affinity purification...biotinylation and purification strategy outlined here provides a basis to determine binding kinetics using a ...

... to improve viral vector production. Whether redesigning viral packaging plasmids, investigating different...untreated and sucrose treated viral preparations.    Despite observing a positive effect on our overall yield...due to impaired endocytosis. In parallel assays designed to test the endocytosis of a fluorescent dye and...

...Addgene is building our antibody resource, which includes recombinant antibodies, educational materials,...California Davis School of Medicine. This deposit includes frequently cited antibodies for use in brain imaging...supporting hybridoma sharing within the community for decades. Educational Resources The Basics of Western Blotting...

...available for binding change as well. If this change hides the epitope that the antibodies used to recognize...conditions. In those cases, pay special attention to designing your controls in validation, as you may need to...help you avoid a great deal of frustration when designing and troubleshooting immunoimaging assays. It can...

...blocks of polymers are heated up and pulled to a desired thickness. The fibers are drawn with microelectronics...whistles’ – including microscale light emitting diodes, thermal sensors, and microfluidic channels. These...using NeuroStack and the fibers as previously described, then measured electrophysiological potential ...

...retrovirus, lentivirus, adenovirus, or AAV. Then, your desired DNA to be integrated can be cloned into the vector...packaged, when the vector is expressed. Once the design and construction steps are complete, it’s time ...weeks, depending on the virus, cell type, and/or desired event.   Figure 1: Transduction to expression...

... mountain – with many routes up and only one destination – a career in the sciences is more like an intergalactic...time working through the details of experimental design and data with authors and referees. And I am constantly...visualization of science? Look at our graphics and design editors. How about condensing a week’s worth of...

...inversions, deletions, and translocations. It also provides researchers with many different ways to control...the right one. In this post, Addgenie Melina Fan guides us through the basics of the validation process...experiment? In this post, Addgenie Chari Cortez guides us through some common controls and why you might...

... the summer of 2013, a remarkable nine papers describing CRISPR/Cas9 genome engineering methods for C....America. To use this SEC-based system, first design and clone 500-700 bp homology arms for your gene...injected 60-80 animals per gene, they obtained the desired insertions on the first attempt for 6/7 genes. ...

...As with other works of Dr. Sacks, Musicophilia includes case studies of relevant neurological atypicalities... Joanne Kamens, Executive Director  I Contain Multitudes by Ed Yong "Microbes are everywhere and this ...shocking at times."     - Jason Snair, Product Designer Sarum: The Novel of England by Edward Rutherfurd...

...distribution of cell lines from repositories on opposite sides of the world shows that science is truly a global... et al., 2006), the hACE2 transgenic mice were designed after it was discovered that SARS-CoV, which entered...dedicated facilities and manpower that a repository provides. Repositories around the world facilitate the ...

...s time, we’ve published an article on BioRxiv describing a comprehensive SARS-CoV-2 virus-human interaction... viral proteins except for Nsp3 and Nsp16, and includes an additional catalytically dead version of the...the viral protease, Nsp5. Our set also includes a GFP construct as a control. For easier portability, ...

...phrase gram negative or positive being used to describe a species of bacteria, but what does it actually...bacteria. In a gram negative bacteria, the alcohol destroys the outer lipid membrane, and the thin inner peptidoglycan... is also useful in a laboratory setting. When designing a plasmid to be inserted into a specific bacterium...

...transcription factor library The human genome encodes approximately 1,800 TFs, with >3,500 unique isoforms... and screening mechanisms, although the unique barcodes for each TF help streamline identification. While...a disease in a model system. TF screens can be designed to identify custom TF programs to acutely initiate...

...abstracts were driven by the quality of the study design or by their preconceived ideas about the controversial...transmission and how different fabrics and mask designs could work to slow the spread of illness. I later...Series: Masters of SciCommSummer Scicomm Series: Modes of CommunicationCareers in Science Communication...

...imaging include ones designed for CFP, YFP, and RFP or the Sedat Quad filter set, designed for DAPI / Fluorescein...brighter signal than mCherry. In general, filter sets designed for Rhodamine / Cy3 will work better with shorter...

... craft sentences for abstracts and presentation slides, learn from audiences as they do science outreach...about whether the same principle applies to other slides in your deck.  If someone asks a question about...of a new science communication initiative that provides yet more scientists with opportunities to practice...

...positive effect on cancer treatment (Ref 1). As described in the presentation, starvation or the use of ...field of Immuno-Oncology, a sizzling hot topic, includes studies on novel cancer therapeutics such as  ... with T cell receptor engineering (Ref 5)  as described by Blue Flame Addgene depositor Professor Michel...

...tables with fluorescent protein properties and provides a quick way to compare fluorescent proteins, e.g...been used in yeast by Lee et al (2013) and in nematodes by El Mouridi et al (2017) and Heppert et al (...practical brightness. Therefore, practical brightness provides a better picture of what can be expected in a ...

...editing!  A recent paper from Feng Zhang's lab describes how to use Cpf1 for multiplex genome editing.  ...sequence NGG TTTV PAM site preserved? Usually Destroyed Yes, Cpf1 cleaves 5' of the protospacer Multiplexing....  Similar to a jigsaw puzzle, the oligos were designed with sticky ends that only anneal together in ...

...immunoprecipitation (ChIP) is an extremely useful technique that provides insight into protein:DNA interactions. ChIP works...up the DNA, or enzymatic digestion. Sonication provides a truly random digestion, but is harsher and requires...both tissue or cell type, amount of protein, and desired fragment length for your output assay. A longer...

...engineering approaches and scientific know-how to design/build/test their own project ideas using synthetic...newsletters, pairs teams with STEM mentors, and provides a nationally recognized platform in March to exhibit...s BioBuilderClub teams’ biological engineering designs and ideas ranged from a biological breathalyzer...

... also easy to run things in parallel. Addgene: Besides being fast and efficient, what other challenges...wanted something that would allow automation. This design cuts out weeks of waiting and hands on labor. That...Addgene, now these can be available to anyone for a modest cost without having to start all over or write ...

...bacterial lac operon contains a gene called lacZ that encodes for the enzyme β-galactosidase. Expression of the...process is called α-complementation. The system described above was put to practical use in the following...Although blue-white screening is probably the most widespread way to select for plasmids containing an insert...

...and does not consider the missing or damaged nucleotides that are common to biological sources of DSBs...better substrates for ligation. Although we do not describe these steps here, the processing of DNA ends tends...of the canonical Cas9 approach. A pair of CRISPR guides that flank regions of hundreds of base pairs or...

...TRIM21 recruits ubiquitin and the proteasome to destroy antibody-bound pathogens. Clift et al. found that...necessary for studying endosomes, the Hell lab designed their super-resolution pH indicators (SRpHi) biosensors...sgRNAs are used with FIRE-Cas9, experiments can be designed to target multiple loci at once. In addition, ...

...transfer origin as well as a selectable marker and a desired genetic payload (ex: GFP). Thus, cells can be both... RNA toehold switch, a RNA hairpin that can be designed to bind and detect nearly any RNA sequence. Downstream...components for gene disruption or CRISPRi. While designed to study the bee microbiome, these broad-host-...

...donated skin, blood and semen samples. Ma et al. designed plasmids containing Cas9 and two different gRNAs... at 3 days post fertilization. Since Ma et al. designed a repair template with two synonymous mutations...thus quieting some fears about the creation of “designer babies.” The differences in editing between iPS...

...ortholog requires a different PAM sequence, a gRNA designed for one dCas9 should be specific to that ortholog...boxB Magenta boxB-MS2 White boxB-MS2-PP7 Despite the success of Ma et al.’s approach, fluorescently...genome editing. Such a system has been previously described for transcriptional activation/repression and ...

...is tested following each immunization until the desired level of antibodies is reached - typically within...within 2-4 months. After the animal has the desired level of antibodies in its bloodstream, the antibodies...species. Pre-adsorbed polyclonal antibodies are designed for experiments where antibodies raised in several...

...serotypes The names of AAVs contain two numerical designations, separated by a slash, that indicate the ITR...interested in expressing multiple small genes, you can design single rAAV vectors with multiple genes packaged... an option (McCarty et al., 2001). This AAV is designed so that the single stranded DNA folds back on ...

...you labeled your gene of interest incorrectly on slides 8-16. Your notes disappear. The wrong version of... speaking is hard. Aside from the diving board charades, I doubt anyone but me remembers any of the incidents... biomedical field, with very little text on the slides and notes instead of written speeches. When you...

... In frogs, ectopic expression of ion channels, despite having an otherwise normal genomic background, ...Subsequent cutting results in a two-headed worm, despite the fact that it’s genetically wild type  (Oveido...fluorescent voltage indicator with near-complete desensitization in continuous light. ArcLight Q239 - A GEVI...

...GFP-labeled cells. Once Tang and Cepko found a description of GFP-binding nanobodies, the project really...stable inside cells. Nanobodies binding GFP were designed in 2009. Subsequent work showed that nanobodies...green-red Cre-lox systems. T-DDOGs can also be designed with other DNA-binding domains, including the ...

...growing dialogue on the state of a system that, despite all of its successes, has many deep-rooted flaws...all bad, however - there are some signs that the tides may be turning. In 2013, the NIH announced its first...in Scientific Training (BEST) awards, which are designed to support new approaches that address the need...

...throughout the library preparation process and includes Picogreen quantification of various intermediate...sequences for Illumina sequencing as well as unique barcodes per sample so we can assign sequencing reads to... use a modified version of the Python program described by Joung et al. 2017, which we hope to make available...