...from Addgene right on our plasmid pages and provide a citation that you can put straight into your reference...our website using the search bar found at the top right of every page. You can search for plasmids by name... number, or keyword. Although an initial search might give you many results, you can narrow your search...search results using the filters found on the right side of the search page. A great breakdown of our search...special collections pages. These pages serve to highlight plasmids that can be used for specific purposes...Every quarter we release a Newsletter where we spotlight some of the plasmids that have come through our...page - each day we showcase a new plasmid that you might otherwise miss. We hope this feature will draw attention...

...Every few months we highlight a subset of the new plasmids, antibodies, and viral preps in the repository...Improved voltage indicator: JEDI-2P Novel class of light-gated potassium channelrhodopsins Myc-tag antibody...indicator (GEVI) called JEDI-2P. Using a custom, high-throughput screening platform to apply saturating...10.1016/j.cell.2022.07.013.   Novel class of light-gated potassium channelrhodopsins by: Mike Lacy ...Xue and John Spudich identifies a novel class of light-gated potassium channelrhodopsins, named Kalium ...2022). The authors show that HcKCR1 and HcKCR2 have high selectivity for K+ over Na+ and rapid photocurrent... optogenetic use.   Fig. 2: A) HcKCR1 is highly selective for K+ over other Na+ and other cations...

...Every few months we highlight some of the new plasmids, antibodies, and viral preps in our repository...different genomic loci — in some cases, over 20-fold higher editing than PEmax. The authors hypothesized that...immunoblotted against vinculin as a loading control (right panel). Image from Addgene. These antibodies... subthreshold voltage fluctuations, while having high fidelity during action potentials of fast-spiking...j.neuron.2023.03.009. PMID: 37015225.  Simple and highly specific targeting of microglia with AAV By Alyssa...human IBA1 promoter (hIBA1a, 466 bp) that drives high microglia-specific gene expression when incorporated...hIBA1a promoters in mice. B) Quantifications showing high microglia specificity of GFP expression for the ...

...upper right) and a mixed population of B cells and dendritic cells (DCs) in the lower middle right. DCs...generated often looks like the graph on the left, it might make more sense once you realize events are plotted...plotted on an invisible Cartesian plane (right.) Each event is plotted by the amount of signal read from...cells. If you look at the dot plot in Figure 2, you might assume that all cell populations are positive for...another two are B+. Our A-B+ cells are in the lower right corner.      Figure 2: This flow plot has... (SSC) and forward scatter (FSC), which measure light that “scatters” past the cell, instead of fluorescent...DCs), mDCs and pDCs. Let's follow it from left to right.  First, SSC and FSC are measured to differentiate...

...FRET signal you should choose the highest quantum yield donor, the highest absorbing acceptor and fluorophores... similar brightness - even though in the most popular pair, CFP has five-fold less brightness than YFP...that case the cross-talk is high and your background signal may be higher than the signal generated by... the acceptor (linking the quantity of absorbed light, at a given wavelength, to the concentration of ...that can affect FRET measurements include: the brightness of a fluorophore pair, donor:acceptor stoichiometry...the donor is excited by a specific wavelength of light and the signal is collected by using emission filters... of the donor is increased. This method is straightforward and quantitative, but it is destructive and...

...insertional effects in the genome. (Bottom panel, right) A highly expressing RUBY plant is a vibrant red/magenta...transparent tissues, or in green tissues is slightly darker or slightly brown. Photos by Andrew Willoughby ...something new has been added to the palette. RUBY is a bright red pigment reporter whose expresion is visible...RUBY as a selection marker The creators of RUBY highlight its use in crop plants under field conditions ... further experiments.     Fig. 3: (Top right) A RUBY-expressing Arabidopsis seed following floral...from the same plant beginning to germinate. (Top right, second panel) Transformed seedlings can be selected...

...listen, and an ability to remember the right stories at the right time. Like many of the posts in our ...be considered in the right, and one in the wrong. If you arbitrate, one party might get exactly what they...I first realized I might have some minor skills in conflict resolution after college, when I discovered...inclination for this skill that you could strengthen. The right set of transferable skills, coupled with your technical...primary role is customer support, but there are eight other teams who still retain a customer support ...be wary of anyone who is actually looking for a fight. But you also can’t be the type of person who avoids...or invalidated. Both compromise and arbitration might remedy short-term substantive conflicts, but they...

...and it emits light that peaks at 508 nm. The protonated state, however, does not absorb light at 488 nm.... efforts have increased the brightness of these genetically encoded light emitters (i.e. Takai, 2015 and... enhanced GFP (EGFP), a variant with improved brightness (Tsien, 1998). Another mutation (A206K) is necessary...of the available green fluorescent proteins emit light. The pKa of fluorescent proteins can be accurately...that are engineered to bind specifically and with high affinity to fluorophores. Examples are the FlAsH...fluorescence overlaps with autofluorescence The cyan light that is used to excite GFP, may also excite several...shifted to longer wavelengths. Therefore, the use of bright red fluorescent proteins (e.g. mScarlet) or infrared...

...illuminates the passing cell with a laser and detects the light scattered by that cell at a low angle (FSC) or large...voltages to observe your cells on a flow plot, you might have to do some testing or ask experienced colleagues..., morphology, and structural complexity.   Height, Width, and Area Within FSC and SSC, there are ...are three parameters that are being measured: height (H), area (A), and width (W). The different measures...few events at the end of your gating pipeline, it might be worth going back and loosening the FSC/SSC gate...Figure 2), single cell events will show up in a straight line one the plot, while duplicates deviate from..., it can depend on the type of stain whether a tighter or looser gate makes sense. Here, luckily, the ...

...the PCR process, you need a high-fidelity polymerase. There are many high fidelty polymerases on the market...Cornell University. Site directed mutagenesis is a highly versatile technique that can be used to introduce...SNPs in a plasmid context). The technique is also highly relevant in this age of CRISPR; site-directed mutagenesis...site-directed genome editing requires a template of high homology to the endogenous target, yet to facilitate...Restriction Fragment Length Polymorphism (RFLP) analysis (right). Introduction of restriction site “A*” results ...rather than a linear product (see figure). Template A high purity plasmid prep significantly increases the ... target site, can identify these duplications (slightly larger bands sizes relative to the original template...

...provides an example calculation of the weights. Table 1: Journal Weighting Example Article Number (A) Year... with Addgene Plasmids in Journal (F) Weight (=F/E) (G) Weighted Citations (=C*G) 1 2010 3 Nature...associated with Addgene plasmid deposits are more highly cited, we went to one of the largest bibliographic...get our journal-adjusted citation measure we re-weight the 3 percent of Nature papers without Addgene ...they comprised 10 percent. Based on this new re-weighting, we can compare citations where an equal share..., you can see the full impact from using these weighted numbers. Note: this does drop some journals from...all other explanations, we do think that it lends weight to the argument that Addgene is achieving its goal...

...complete desensitization when exposed to continuous bright light. Desensitization is mediated by a conserved ...Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids...MerMAIDs, neurons can be turned off quickly and with high precision - a valuable optogenetics feature when...PubMed PMID: 31346176. Using LADL to loop with light Article contributed by Eric Perkins Mammalian genomic...looping, the Phillips-Cremins lab developed the light-activated-dynamic-looping (LADL) system. The group...protein that heterodimerizes with CIBN upon blue light exposure (see Fig 1*). When the group targeted the...increased Zfp462 expression within 4 hours of blue light induction (6-fold faster than previously developed...

...grant. Here’s a list of tools and resources that might help make writing a little easier. Writing schedule...when writing for a general audience. The tool highlights jargon terms or words rarely used in general ...frequency of use in orange, and common words with high frequency of use in black. This paper talks more...either based on the section of a manuscript they might be found, i.e. introduction, methods, results; or...Mendeley is free to use. You can take notes and highlight the papers saved to your Mendeley library. The...have a lot of the same features, but this chart highlights the differences between these two citation managers...

...the left while the SnapGene powered map is on the right (click here to see the new map in action). Read ...Analyze your plasmid sequence For more immediate, high-level analysis, you can click directly on the “Analyze...number of times they cut. Use the filters on the right to find single, dual, and 3+ cutters. You can also... on a particular restriction enzyme here also highlights it in the “Map” and “Sequence” tabs. Features...types. Clicking on a particular feature here also highlights that feature in the “Map” and “Sequence” tabs...direction. Clicking on a particular primer here also highlights it in the “Map” and “Sequence” tabs. BLAST...

...Mutagenesis) High Gene Expression Infects Non-Dividing Cells Large Packaging Capacity (>8kb) High Transduction... it encodes. This ability to infect cells with a high efficiency and deposit a DNA/RNA cargo into their...than the alternatives? Well, for one they have a high transduction efficiency and can effectively infect...organisms. Finally, viral delivery is typically straightforward across applications, while electroporation ...upcoming post. If a viral vector seems like the right approach for your experiment, you’ll need to decide...packaging is less efficient when inserts are on the higher end. Adeno and adeno-associated viruses Adenoviruses...do. They are generally used to achieve transient, high expression of the relevant cargo without the fear...

...and on-targeting likely more efficient.  These insights are hard to appreciate without computational support...must be confusing for a newcomer to decide on the right software to use. With so much choice the question...something akin to a frenzy as the crowd tries to weigh and determine the value of every sweet or toy strewn... audience blindly receiving them; it results in highly similar content and the absence of iterative design...result, the software piñata has been filled with slight variations on a theme without many resources to...tools The goal of this post is to provide some insight into available CRISPR software tools, what problems...various tools are trying to solve, and finally how we might proceed in the future. When we think of designing...

...two appear on an agarose gel at notably higher molecular weights than expected, given the size of the plasmid...plasmid can exist on as a dimer, depicted on the right, with the entire monomeric sequence duplicated in...), direct repeats (Ribeiro et al., 2009), and at high copy numbers (Williams, Carnes, and Hodgson, 2009... monomers, they are also able to replicate at a higher frequency (Summers & Sherratt, 1984; Summers et...are larger than monomers and will therefore run higher on an agarose gel (Bedbrook & Ausubel, 1976). In...2011) indicated that strain HB101 produced the highest yield of monomeric plasmid compared to DH5α. However...: 7029308. Goebel W, Helinski DR. Generation of higher multiple circular DNA forms in bacteria. Proc Natl...

... standard antibody: An ~150kD protein with both light and heavy chains made by immune systems. But here...imaging applications, smaller affinity reagents means higher resolution. Many of the below are also small enough...the lab, instead of generated in vivo, they have higher reproducibility and consistency in the final product...cleavage. They are comprised of the variable heavy and light chain regions, connected by a short linker peptide... longevity compared to antibodies, along with a higher likelihood of aggregation (Bates, 2019). They are..., are derived from a camelid antibody lacking a light chain, called a heavy chain antibody (Muyldermans.... DARPins typically consist of 3-4 repeats, have high specificity and binding affinity, can be produced...

...supervisor might expect weekly or biweekly progress reports. However, a hands-off busy supervisor might get ...Know-it all experts, who think their way is the only right way, and Indecisives, who keep changing their ... your discussion will get side-tracked, and you might create even more conflicts. 4) Define in advance...about mutually beneficial solutions Your supervisor might not agree with all of your ideas, but he or she ... conflict is miscommunication. For example, you might misunderstand your supervisor’s suggestions and ...sense their tone of voice in an email, there is a high likelihood of miscommunication, which can lead to...assertive to get your point across. Some supervisors might try to intimidate you with negative comments (hostile-aggressives...

...technologies, one might incorrectly relegate RNAi to history. The straightforwardness of RNAi is unmatched—unlike...considered the “gold-standard” for LOF studies, the high gene copy number found in plants makes traditional... DNA, thereby limiting CRISPR’s effectiveness at high copy numbers and genomic loci with low accessibility...directly compared CRISPR to RNAi in the context of a high throughput survival screen found that either technology...for cell growth. Together, this excellent work highlights the importance of using orthogonal methods to...Arguably the greatest advantage of RNAi is the straightforward nature of the technique—all that is required... long hpRNA with Gateway-based cloning and straightforward positive selection of plants undergoing RNAi...