...science in D.C. "Scientists shouldn't be hidden figures.  Ask me about my science!", read my sign.  People...University. She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207...data collection, destroys existing data, and guts future funding.I marched because science belongs to the... some little scientists (as you can see in the picture) and this march proved to be the perfect opportunity... a Scientific Instrument Maker at Intellectual Ventures Laboratory. Vancouver, BC Canada 49.2827°N 123.1207...attended the march in Vancouver, BC, although I wasn’t sure I’d go until several hours before it started. It...Previow Post on the March for Science Learn How Future of Research is Advocating for Change Within the...

...FPs to highlight various subcellular structures. Please see Figure 1 for commonly used markers for major...up of organs, cells comprise compartments and structures, called organelles. Take a sneak peak inside ...fluorescent protein fusions to visualize subcellular structures. For example, you might use your fluoresent protein...organelle of interest. Click on the gene names in the figure below to find plasmids containing that particular...find more plasmids for labeling your subcellular structure of interest in the Allen Institute for Cell Science...possible to get an idea about the subcellular structures your protein resides in and potentially which...proximity of two proteins within the same subcellular structure or protein complex. By coexpressing both the marker...

...with the literature There are a few ways to make sure you are keeping up with the literature in your field... with the scientific literature a little bit at a time. Searching for literature My go-to search engines...the literature until you have all of the information you need. Reading primary research literature The...papers are also just poorly written. By nature, scientific literature is information dense and since it continually...you daily updates with relevant literature. Organizing the literature As a scientist, you will begin to...require numerous other references to scientific literature to substantiate the facts upon which it builds...means you have to become familiar with a body of literature related to the topic. Finding reliable references...

... manipulate biology based on molecular features found in nature, with the ultimate goal to improve the...impact on this challenge either for magnetic capture or for capture using microfluidic devices. Addgene: What...antibodies against common tumor markers to subsequently capture cancerous cells expressing low levels of tumor ...binding stability really mattered for efficient capture. We wanted to be able to build a highly multivalent...for keeping going while we worked out the cell capture system.) Cancer and other applications Addgene:...technology mean in the context of cancer? Howarth: The capture of circulating tumor cells (CTCs) is going to have...treatment of cancer. It is clear that one needs to capture a diverse range of CTCs to cope with the variation...

...addition of a structured motif to enhance pegRNA stability PEmax - optimized PE architecture; increased ...synthesized (Figure 8A), then cloned in a pooled format into lentiviral transfer vectors (Figure 8B). Libraries...Berkeley. Plan Your CRISPR Experiment Resources Figures All figures were created using BioRender.com (Link opens..., N., Ishitani, R., Zhang, F., & Nureki, O. (2014). Crystal Structure of Cas9 in Complex with Guide RNA...vivo genome editing using Staphylococcus aureus Cas9. Nature , 520 (7546), 186–191. PMID: 25830891 Rauch...became the most popular genome engineering approach. Figure 1: Overview of the basic CRISPR mechanism Engineered...gRNA left free to interact with the target DNA. Figure 2: Overview of the NHEJ repair mechanism Cas9 will...

...isolated and placed in a buffer. Antibody Structure Figure 1: Structure of an antibody A standard antibody is...Antibodies can also be produced in cell cultures using hybridomas, cell cultures created by fusing B cells with...form a “Y” shape, as shown in Figure 1. The two arms of the Y structure are responsible for binding the...Monoclonal antibodies are generated from cultured B cells, where each culture should produce the same antibody...Diabodies can be used for assembling protein nanostructures. Figure 2: An antibody (IgG, far left) compared... a signaling molecule. Figure 4: A direct sandwich ELISA, which uses a capture antibody and a primary ...In a sandwich ELISA, a capture antibody, bound to a surface, is used to capture the protein of interest...

...Guide to Channelrhodopsin Variants: Features, Limitations and Future Developments. Exp Physiol. 96(1): ...optogenetics integrates optics and genetic engineering to measure and manipulate cells (frequently neurons) and their...optogenetics research utilize light to detect, measure, and control molecular signals and cells in order...functions. Example schematic of an optogenetics procedure. A channelrhodopsin, fused to mCherry, is expressed... allow the fast depolarization of neurons upon exposure to light through direct stimulation of ion channels... in excitation (depolarization) of the neuron. Feature enhancements include: Increased photocurrent amplitude...from the cryptophyte Guillardia theta ). Other feature enhancements include: Increased photocurrent amplitude...

...interest is PCR amplified, PCR products are denatured-renatured, treated with a nuclease that cleaves DNA...reagents are appropriate for a given experiment. Make sure to check whether reagents are available to carry... gRNA In vitro transcription reactions generate mature Cas9 mRNA and gRNA, which are then delivered to...match the target locus, but it is also critical to ensure that the gRNA target sequence does NOT match additional...allele may be different owing to the error-prone nature of NHEJ. In HDR gene editing experiments, most ...protein-based genome editing in vitro and in vivo. Nature Biotechnology . 33 (1), 73–80. PMID: 25357182...

...pSico plasmid to drive shRNA expression. Other features that can be included in transfer plasmids include...contains the viral LTRs and psi packaging signal (not pictured). Unless an internal promoter is provided, gene...Plasmids 2nd vs 3rd Generation Lentiviral Systems Feature 2nd Generation 3rd Generation Transfer Plasmid ...Invitrogen Stbl3™ or NEB Stable cells. This will ensure that the repeats will be maintained and often results...always be considered with respect to the precise nature of experiments being performed, and your biosafety... Genetic Organization and Genetic & Structural Features . In vivo gene delivery and stable transduction...

...check out our Chemogenetics Plasmid Collection . Figure 1: General overview of a chemogenetics experiment...muscarinic receptor and has not been used in vivo . Figure 2: DREADDs, their ligands, and signaling properties...and/or K + ions, and activates neuronal activity. Figure 3: PSAM4-based ligand-gated ion channels, their...optogenetic systems in our Optogenetics Guide . Figure 4: Luminopsins can be activated through chemogenetic... control expression of a chemogenetic reporter ensures that the reporter remains silent until a cell or... X, Stone O, Roth BL, Jin J (2015). The First Structure–Activity Relationship Studies for Designer Receptors...

...propagation of these plasmids through cell division. To ensure the retention of plasmid DNA in bacterial populations.... Under these conditions, there is a selective pressure to retain the plasmid DNA, despite the added replication...organisms, including plants, worms, mice, and even cultured human cells. Although plasmids were originally...bacteria, but also provide those bacteria with a pressure to maintain and replicate your plasmid over multiple... extremely important when verifying plasmids to ensure each plasmid contains the essential elements to...replicated using the modified bases and a microscope captures the fluorescent color that is emitted each time...

...plasmid; the separation of Rep and Cap shown in the figure on the right facilitates the viral pseudotyping...genome. This is important because local chromatin structure at the site of genome integration can affect the...optimized for use in AAV, or containing Staphylococcus aureus (SaCas9) . AAV is also a promising method for gene...or a genotoxic agent such as UV radiation or hydroxyurea, is necessary for lytic cycle activation. When...concatemerize, producing high molecular weight structures that are maintained extrachromosomally. rAAV ...

...conjugated to a signaling molecule, can be used to capture, quantify, and identify proteins of interest. Read...Optogenetics integrates optics and genetic engineering to measure and manipulate cells (frequently neurons) and their...optogenetics research utilize light to detect, measure, and control molecular signals and cells in order...

...When you're not sure where to start or need a handy reference, Addgene's eBooks are here to help!...Addgene's eBook Collection When you're not sure where to start or need a handy reference, Addgene's eBooks...you need to know about plasmids, from parts, nomenclature, and uses to common promoters and cloning techniques...

...infected cell receives only one plasmid. To make sure that every plasmid is adequately represented in ...planning to conduct a pooled library screen. The figure above is an outline of how a gRNA library is used...cells that survive post-selection. The selective pressure must be strong enough that most of the cells die...

...the nucleus through nuclear pore complex. This feature may be useful for the transduction of terminally...-retroviral plasmids have similar names? The nomenclature of the plasmid backbone is meant to be descriptive...

...the TSS. Eukaryotic promoters are so complex in structure that the DNA tends to fold back on itself which...which is a main component of a cell’s ribosome structure. Ribosomes are the site of protein syntehsis where...

...sequencing primer" and "3' sequencing primer". Still not sure what primer you need? Email us at [email protected]...

...protocol on our website. Ligation Independent Cloning (Figure adapted from LIC protocol ) Back to Top Yeast-mediated...

...crystallography cannot capture flexible or disordered regions, so crystal structures like those in Figures 2 and 3 don...biology! Major features First, we’ll cover the basic parts of the Cas9 enzyme.   Figure 1: A cartoon...from Streptococcus pyogenes.   Figure 2:  Crystal structure of S. pyogenes Cas9 in the apo state...state that it doesn’t appear in the crystal structure (Figure 2) at all. When this crucial recognition region...learn about this process!      Figure 3: Crystal structure of S. pyogenes Cas9 bound to guide RNA...are disordered and don’t appear in the Figure 3 crystal structure, reflecting the less stringent recognition...tolerate mismatches in this region. The crystal structure in Figure 3 shows the guide RNA stabilizing the DNA-binding...