...(Cetn1/sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence indicates a more successful...you can hold off on genotyping until the pups are 2-4 weeks old and use the ear tag method (creating different...PMID: 24253446. PubMed Central PMCID: PMC3875854. 2. Mashiko, Daisuke, et al. "Generation of mutant mice...plasmid using standard cloning techniques (The XX indicates where your target is inserted, disrupting the ...effective ones to increase your chances of gene modification in vivo. Find Plasmids for Testing Your gRNAs...et al., 2014). The plasmid is sturdier than the delicate RNA and therefore there has less chance of contamination... the validation assay should give an accurate indication of how many pups will contain a mutation and ...

...under phase contrast starting day 2 after transfection.     Fig: 2: Expressing mARGAna in MDA-MB-231...engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase and outgrow the... they are based on light which is very easily scattered by tissue. Ultrasound, on the other hand, is a...protein nanostructures called gas vesicles, which scatter sound waves and thereby produce ultrasound contrast...limiting their applicability. They also did not scatter ultrasound nonlinearly, making them difficult to...out to develop next-generation bARGs that could scatter ultrasound nonlinearly and be easily expressed ...promoter pBAD and a low-copy number origin of replication (p15A) worked best under a wide range of inducer...

...presented in a series of quick (10 minute) or very quick (2 minute) talks so we got to hear from everyone. Addgene...evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (Andrea Sweigart) Monkeyflowers...Organisms, Part 1 Five Popular Model Organisms, Part 2 ... their organismal systems for scientists and educational programs for K-12 students. You can see why I...parenting strategies and make important ecological indicators since species live in water and on land. Current...which have been used to treat heart disease. Caterpillars that eat milkweed generate cardenolides that... poisonous) for bird predators - birds eat a caterpillar once, vomit, and never do it again. Cichlids ...

...improve solubility and stability (Figure 2).      Figure 2: Crystal structures of CRBNmidi alone...fluorescent tags in Caenorhabditis elegans. Genetics, 228(2), iyae126. https://doi.org/10.1093/genetics/iyae126...performs well in many different antibody-based applications.  IPI’s recombinant version is a fusion of mouse...with additional cofactors or using significantly truncated forms that limit its functionality. To make biophysical...cellular processes in C. elegans: mScarlet3 for applications needing intense brightness (such as low-expression...luciferase fragment (expressed separately) assemble and catalyze the luminescence reaction (Figure 5). The researchers...under tension. Upon binding, LgBiT and BiT Tag catalyze a luminescent reaction with the substrate furimazine...

... Figure 2: Ironing boards. Good as a WFH desk, and as a video control center.   Cycle 2: Zooming ...ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple, external video...Creating great educational science videos is tough. Is the content accurate? Are we conveying complex... had set up involved multiple people in tight locations. Conference rooms for on-screen presenters! Labs...wasn’t going to work for the majority of our educational videos. In addition, it’s really asking a lot...receiver, to beam the camera’s image to my remote location, live. After doing a bit of looking, I found an...

...identifying the resources needed to accomplish that goal. 2. Identify the tools in that timeline Avoid writing...based on experimental timelines and specific aims. 2. Then make decisions based on financial values. All...Would I be able to get access to these materials?” 2. Always negotiate for a win-win decision An institute.... After identifying all the major equipment, categorize all of your minor tools, like pipets, or mini-centrifuges...titles in the regions where the host institute is located. For example, salaries in Kansas versus salaries... most start-up P.I.’s. It doesn’t have to be complicated, or difficult to create this list. Start BEFORE...discounts, collaborations, access to internal award applications, etc. If this is the institute that can help...

...aspects of the work you presented may also be unclear. 2. Make the audience your primary focus When writing... the Communicating Science Conference series for graduate students. I believe that communication is the...skill in science communication over time.  Like most skills, great science communication comes with practice...able to communicate with their colleagues and peers. Scientists also need to be able to communicate to expert...need to communicate with non-scientists, to establish broader impacts for their work in education and policy...with opportunities to practice communicating.  Some fantastic K12 education initiatives like BiteScis (https...careers in science communication The strength of story telling Science communication: 9 strategies to get...

...perceived climate change risks. Nature Climate Change, 2(10), 732-735. doi:10.1038/nclimate1547 Kunda, Z. (...Further, when science is communicated in a format that is accessible, that communication often neglects the ...SciCommSummer Scicomm Series: Modes of CommunicationCareers in Science Communication: Science Writing... to save a few days’ worth of capsules to self-medicate with when they get a cold. A primary school teacher...dihydrogen monoxide (H2O). Studying science communication as a PhD student Fortunately, by the time I ...Otago had established the Centre for Science communication and I was accepted as a grad student. I chose...a phenomenon called “motivated reasoning,” the catchall term for a range of neat little tricks our minds...

...highly efficient for C. elegans genome modification. Figure 2: Schematic from Dickinson et al. (2015...one injection step! The entire protocol takes about 2-3 weeks but requires less than eight hours worth of...genomic modifications, is very robust in C. elegans, and the methods for HDR-mediated modification continue...After self-excision, the resulting loxP scar is located in this synthetic intron, and only the fluorescent...permission from the Genetics Society of America. Applications of this system Dickinson et al. have developed...system could be used to make many other genome modifications, including targeted mutations. SEC-based systems... in cell culture. In addition to CRISPR/Cas9 applications, SEC selection could replace other, more restrictive...

...native enzyme, which are summarized in Table 2 below.  Table 2: Properties of native luciferases Organism...Luciferases are a class of enzymes capable of catalyzing chemical reactions in living organisms resulting...gene, such as luciferase, usually serves as an indicator of transcription within cells, where detection...photon emission is measured resulting from the catalysis of a chemical reaction requiring luciferin, ATP...Background Lower Higher Post-translational Modification Not Required Required Photobleaching/Phototoxicity...blue light with a peak emission at 480nm upon catalyzing a chemical reaction with the substrate coelenterazine...compatible substrate must be provided for the enzyme to catalyze the light-producing reaction. Many luciferase ...

...phages but to also destroy the phage upon re-exposure (2, 3). More specifically, specialized Cas proteins snip..., accessed 25 November 2013. Author: James Atmos (2). Many CRISPR/Cas moieties carry out these functions...distinct types and subtypes using polythetic classification based on evolutionary similarity (5). Type ...2013 (7, 8). Nine months later, more than 1500 publications describe work to improve the tool’s specificity...various species and describe an assortment of applications. Despite the ubiquity of CRISPR/Cas toolkit,... multiple plasmids used for various platform applications. For more information on CRISPR/Cas, check out...for your research. Do you have a cool idea or application that involves CRISPR/Cas technology? Let us know...

...the reporter mRNA is labeled by PCP-mCherry (Figure 2). The 3’ UTR also contains a CAAX sequence to bind...PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor... While ribosome profiling co-opts the physical location of the ribosome, TRICK (Translating RNA Imaging...development, the osk-TRICK reporter was double labeled, indicating translational repression. During later stages...ribosomes after initiation, to calculate a ribosome translocation rate of ~3.5 codons/second. They also explored...Resources the Addgene Blog Learn More about SunTag Catch up on Your GFP Background Choose the Brightest Fluorescent...

...episode 2 "Organizing Literature" : I found out about Zotero here Dear Grad Student podcast episode 2 "Essential...research papers or dissertations. Phrases are categorized either based on the section of a manuscript they...introduction, methods, results; or phrases are categorized by the goal of the phrase, such as being cautious...

...target sites for editing. PAM sequences can be from 2-6 base pairs in length and follow the DNA sequence... Eric Perkins Mammalian genomic DNA is not just a 2-D linear molecule snaking through our cells’ nuclei...plasmids at Addgene Opperman J et al., Nature Communications, 2019. PubMed PMID: 31346176. Using LADL to...light-activated-dynamic-looping (LADL) system. The group fused dCas9 to a truncated CIB1 protein from Arabidopsis (CIBN), which they...

...ITR, which can either be “flip” or “flop” (Figure 2). Additionally, the ITR contains a 4-nucleotide Rep...site for Rep proteins (Savvy et al. 2013). Figure 2: AAV2 wild type ITR in flip and flop orientation. ... ITRs serve as the origin of replication and are required for replication, packaging, and vector persistence...viral rep (replication) and cap (capsid) proteins, which bind to the ITRs and initiate replication. ITRs are...structures, ITRs are prone to replication errors and disruptive mutations and truncations that can make vector ...binding site for Rep78 and Rep68 to initiate replication and a terminal resolution site that serves as...are noted. Created with biorender.com. ITR modifications and types  Since ITRs are one of the most important...

.... Figure 2: As the cell passes through the laser beam and causes it to scatter (FSC/SSC), the detected... Figure 2), single cell events will show up in a straight line one the plot, while duplicates deviate ... intensity of the cell’s illumination (see Figure 2). H describes the maximum signal strength, while W... you with two measurements: forward scatter (FSC) and side scatter (SSC). To acquire these measurements...exclude all duplicate events (two cells stuck together) from your single cells, as duplicates can’t be reliably...types of scattering that occur when a cell is pulsed by the laser. Within each type of scattering, you also...to keep it short though, promise! FSC and SSC: scattering To digitally extract your cells based on their...

...cellular responses in addition to antibody response 2) very high immunogenicity 3) and long-lasting immune...their use in other clinical applications. In the last decade, viral applications in cancer treatment and ...s unpack the myriad number of emerging viral applications and give you some recommendations for putting...putting viruses to work. Viral applications Viruses work by infecting a host cell (the target of viral infection...expressed. It’s important to note that for all the applications we discuss, there is more than one type of virus...retrovirus, AAV, etc.). When you’ve decided on an application, our Viruses as Biological Tools blog can help...viruses and pick one for your experiment. Research applications Gene expression control Researchers are always...

...  AAV9, AAVrg pAAV-hSyn-DIO-mCherry 37825 AAV1, 2, 5, 8, 9 (20 µL size serotype testing) pAAV-CAG-GFP...size serotype testing) pAAV-CAG-GFP 50465 AAV1, 2, 5, rg (20 µL size serotype testing) pAAV-hSyn-EGFP...availability date or to be notified when a particular catalog item is available for order please email [email protected]...

...2013) In Vivo Applications of Single Chain Fv (Variable Domain) (scFv) Fragments. Antibodies 2:193–208 . ...panning and scFv characterization.    Figure 2: Phage display starts with a pooled library of scFvs...the short epitope GCN4, also called a scaffold; and 2) a GCN4 binding scFv that’s fused to GFP. When the...led to their application for basic and translational research.  Basic research applications of scFvs: ...blood clearance, which is useful for imaging applications; better tissue penetration which is useful for... for therapeutic and imaging applications; and reduced immunogenicity when administered in vivo due to...from blood can be a drawback for therapeutic applications where longer retention times often increase ...

...standard mutation rate by only ~2-fold over nCas9 levels. Two sets of modifications were also made to PolI3M ...10-3 mutations per nucleotide per generation, and (2) the addition of the thioredoxin-binding domain (TBD...potential applications. Challenges of existing mutagenesis methods Several continuous diversification methods... challenging for users to achieve targeted diversification of all nucleotides within a defined region ...Chang Lab have created methods that introduce diversification by targeting a site of interest with dCas9 ...level of variation that can be introduced.   Diversification is limited due to the integration of oligonucleotide...increasing the standard mutation rate seen during DNA replication by 120-fold over background. Due to its modular...