...injection site is straightforward, as it targets large muscle groups (usually the thigh), though it is important...injection, like you might after getting a vaccine. Restraint: mechanical restraint Location: thigh Gauge: 25–27...use depends on a number of factors. You have to weigh the pros and cons of various methods and choose ...circulation is slower, and more institutional oversight is required. Restraint: mechanical restraint or...such as experimental therapies or pain relief, straight to the stomach. This technique involves administering...Intermediate Intramuscular Mechanical restrainer Thigh 25–27 < 0.02–0.05 mL Beginner Subcutaneous Back...

...emission spectra, meaning they will emit fluorescent light at similar wavelengths (See this handy emission ...against APC-Cy7, the populations are in more of a straight line and are only positive for APC (Figure 1B)...horizontal line, so our circled population on the right needs to shift towards the X axis to become a perpendicular... plot in 2A, the populations are not perfectly straight, but the center of each population is in a line... sample you are trying to compensate in the top right corner of a flow plot, meaning that it should be..., it will show up as a diagonal line in the top right corner of the plot and large compensation values...positive sample. The cell population in the top right corner of the plot is positive for both markers,...

...for fusion proteins! Despite multiple studies in high profile journal articles, many researchers remain...primarily as a dimer in cells while a protein with a high micromolar or low millimolar Kd is unlikely to form... Following the simple logic outlined above, you might think EGFP is unlikely to form dimers in a cell....then their effective concentration will be much higher than if they are homogenously distributed and mobile...concentration of FPs around that fusion protein is very high. Real world examples of these situations in cells... the POI. These structures are large (microns), bright, non-physiological, and difficult to ignore [1,...normally self-associate into homodimers or even higher order oligomers. Unfortunately, FP dimerization...

...and understanding of CRISPR, and perhaps provide insights and ideas for your own research! Classification...categories, arriving at a consensus was not as straightforward as it seems. There is not a single gene that... sequence similarity, phylogenetic analysis, neighborhood analysis of surrounding genes, and experimental...cutting up DNA after recruitment by Cascade. As you might imagine, this is handy for creating large genomic...adaptations for increased fidelity, activation/repression, high precision edits, and more. Figure 3: Schematic...Cas12 create short 3’ overhangs, which leads to slightly increased efficiency when conducting edits using...with BioRender.com.   What’s your type? It is highly unlikely that you will have to know and understand...

... completed with the Ig light chains. The variable region of the heavy and light chain determine what antigen...going to spot the whole, long word right off the bat, but you might see a few letters and be able to guess...Two immunoglobulin (Ig) heavy chains and two Ig light chains make up each antibody molecule. The Fc region... of an antibody including Fc, Fab, heavy chain, light chain, constant region, variable region   The...made up of the variable regions of the heavy and light chains fused together to form a single protein that.... The proteins are separated based on molecular weight by SDS-PAGE, and are subsequently transferred to...interest and detect its presence. Then, the amount of light that can pass through each well - the optical density...

...accurate and reproducible. I also highly recommend spending the money on a high quality stereotaxic frame with...and versatility, the technique is remarkably straightforward to learn and relatively cheap (compared to ...manipulated a number of the opsin genes to alter the light spectrum for activation and the kinetics of opsin...above) include Channelrhodopsin2 (Chr2), a blue-light sensitive non-specific cation channel, activation...potential. Halorhodopsins (HR) are green/yellow-light sensitive chloride ion pumps, activation of which...expression alone is extremely limited. Therefore, highly expressing, generic neuronal promoters such as ...privately communicated to me as one of these extremely high expressing promoters). Conduct a time course experiment...

...Addgene we've set out to fill this gap by both highlighting that researchers do learn MANY transferable ...the lab and by offering advice on areas where you might need some help. Today in our transferable skills...with SnapGene, for example, allows us to display high-quality maps and annotations without us having to...events can be very tough. First I needed to find the right contacts at the companies and then convince the ...Great progress! You are collaborating with your neighboring lab, you've met interesting people at conferences...are the individuals that are part of them. I’m delighted that these collaborations give me opportunities...An Imperative for Graduate Students" from Inside Higher Ed Additional Resources on the Addgene Blog Check...

...have limited use in vivo because they are based on light which is very easily scattered by tissue. Ultrasound...modality with good tissue penetration depth as well as high spatial and temporal resolution, making it ideal...vesicle genes from Serratia sp. 39006 produced the highest levels of nonlinear acoustic contrast in E. coli...). Gas vesicle expression in vivo is also straightforward: after the E. coli have colonized tumors, inject...Methods to assess bARGSer expression, from left to right: colony whiteness on inducer plates, white inclusions...We wanted to create second-generation mARGs with higher expression, a smaller genetic footprint, and nonlinear...chromatin-opening agents. It ended up being very straightforward to engineer various cell lines from different...

...and writing are key skills. Other fields, often highly technical or skills-based ones, may not use them...are less likely to require cover letters, while higher-level jobs are more likely to consider cover letters...that asks for strong organizational skills. You might think, “I’d be good at that! I organized my lab ...]. Notice that the intros above allow you to highlight what specific skills, qualities, or interests ...considered cover letters to be more important for higher-level positions. Many did not require cover letters...-neutral greeting like “Dear Hiring Manager.” Highlight your achievements with specific examples. Instead...

...parts of our code, we might be calling something a sample and in other places, we might be calling it an aliquot...soundness of the software design. Each of us brings slightly different cognitive biases as a result of our ...creates trust amongst the team. Whether we are a highly collaborative team because we are using the Scrum...we are using the Scrum framework because we are highly collaborative, we love teamwork. Working collaboratively...parts of the shipping code. The following sprint we might have an EMTA theme and we will all be working in... Most of the time, updating these libraries is straight forward. However, there are times where a library...no further updates, security or otherwise. This might sound like Greek to you, which brings me to my next...

...densities that are low enough to prevent confluency but high enough to promote cell growth. Overgrown cultures... age of your culture post thaw. In general, the higher the passage number of a culture the lower the transfection...be a bit dubious due to lack of documentation and high turn-over of lab personnel. If you are unsure of...with pH changes and resource consumption due to the high cell density of the culture. In addition to benefitting...harvest approach is that it tends to result in a higher volume of lower titer virus. In such cases you ...downstream applications. While this ensures that the highest titer virus is used, it is not always feasible,...titered. Most titering approaches transduce into highly permissive cell lines such as HEK293, H0 or A549...

...IBEX is a highly multiplexed tissue imaging method that uses iterative staining and chemical bleaching...bleaching to allow for high-resolution imaging without degrading the tissue (Radtke et al., 2020, 2022). I do... do mean highly multiplexed: you can image over 65 different parameters using this method (see Movie S8..., you’re probably also very interested in having high-quality antibodies and reagents that you know work...you’re certainly not alone – we’re all looking for high-quality antibodies and reagents that work for our...immunolabeling and chemical bleaching method for high-content imaging of diverse tissues. Nature Protocols... Data with Addgene Antibodies 101: Choosing the Right Isotype...

... for a reply to an email. You need answers: the right ones, quickly, and this is why we have a dedicated...phone call or write out an email. You need help right at the moment you’re looking for it, or you may ... from your colleagues, to intellectual property rights and handling your constructs once they reach you... new feature on our website; towards the bottom right corner of each page there’s a blue icon displaying...result. By clicking on the result links themselves right from within the widget, you’ll navigate to articles...on the "Leave us a message" button shown to the right, you will be be prompted to enter your name and ...team’s attention, where we can then offer the same high level of direct service you’ve come to expect from...

...together DNA fragments. As a fun way to start, I highly recommend watching this entertaining video created...assembly work? The required homology between neighboring fragments can be created via PCR amplification...including regions of homology to the vector or neighboring fragment. Then you would amplify the fragments...success rate of this reaction is typically fairly high, so there is usually no need to screen a large amount...structure, such as a hairpin or a stem loop (as might be anticipated to occur within a terminator sequence...required single-stranded annealing and priming of neighboring assembly fragments. Gibson assembly is commonly...vector that you want to use is very large, has a high GC content, contains a lot of repeats--any of which...

...via gene therapy, so researchers have set their sights on another approach. Since shorter forms of dystrophin...to persist over time, stem cell editing would be highly desirable. Should stem cell editing occur, the ...and AAV9, respectively. Although each study used slightly different methods, they each observed the same..., the relative abundance of exon-skipped mRNA is high, likely because this mRNA is not subject to nonsense-mediated... very low to no off-target activity at the ten highest-ranking predicted off-target sites. Each paper ...CRISPR delivery must be optimized to: 1. Reach a high percentage of muscle cells throughout the body, ... rigorously examined over a long period of time. High fidelity Cas9s such as eSpCas9 and SpCas9-HF should...

...transcribe the extremely long, highly structured HCV genome (~9.6 kb of highly structured template!). MarathonRT...Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids...Listen to the Hot Plasmids Podcast!  MarathonRT: A highly processive reverse transcriptase Article contributed...complex heterogeneous RNA samples. Furthermore, such high processivity enzymes and the structural understanding...antibodies: they are small (12-15 kDa), stable, highly soluble, and non-immunogenic yet are still just...

...Syndrome One of those teams is led by Claudio Aguilar (right) at Purdue University and is working to understand...forward. Xeroderma Pigmentosum Hendrik De Buhr (top right) and his colleagues at the Wistar Institute in Philadelphia...condition leaves patients incredibly sensitive to sunlight and prone to developing metastatic melanomas and... and other cancers. Joshua Wang (bottom right), also of the Wistar Institute, says that the lab got interested...Alternating Hemiplegia of Childhood Melodi McSweeney (right), a graduate student at Duke who has recently moved...returning to basal conditions normally after periods of high activity. The new work will help them to understand... Syndrome Last but not least, Meng-Shin Shiao (right) of Mahidol University’s Ramathibodi Hospital in...

...tricky at best, and downright infuriating at worst. While it is tempting to just dive right in and start injecting...also transduce postsynaptic cells when used at very high titers (Zingg et al., 2017), which can lead to cells... total injection volume. You’ll often hear that “high titer” is required for efficient AAV transduction...the spread of the virus.  A better way to define high titer, then, is this: It’s the optimal number of...inversely related to transduction efficacy: the higher the efficiency, the lower the titer and vice versa...needed volume (de Backer et al., 2010).  Second, high efficiency transduction coupled with a strong promoter...amount that looks effective. Ultimately, the perfect “high titer” is one that optimizes gene expression where...

...copies can be tightly controlled by titrating the transposase to transposon ratios. High ratios of transposase...efficiency of excision with PBx has been shown to be higher than that of the wildtype piggyBac™ transposase...importantly, unlike Cre recombinase, the PBx is highly specific, and has not been shown to cause genomic...specialized knowledge to help researchers identify the right technologies and strategies for their projects. ... 2016. PubMed PMID: 27075976. 4.  X. Li, E. R. Burnight, A. L. Cooney, N. Malani, T. Brady, J. D. Sander.... Qualls, B. M. Lewis, E. Ke, C. T. Dargitz, R. Wright, A. Khanna, F. H. Gage, and I. M. Verma, “Functional..., M. van Lohuizen, A. Berns, and D. J. Adams, “A high-throughput splinkerette-PCR method for the isolation...

... in Paris and New York. We want YOU to join the fight against coronavirus We are just at the beginning...be the next scientist joining in this worldwide fight against SARS-CoV-2. As we’ve all been affected in...that we have to stick together (6 feet apart), to fight the new Coronavirus. To help get you started, here... sites - so cloning projects should be fun and straight-forward. All constructs are tagged with a 2xStrep-tag... gave a lot of background in our study - so you might want to be careful with these clones (and help us... research. For all these reasons, the QCRG is delighted to be partnering with Addgene, who will be taking...bloggers Manon Eckhardt (left) and Melanie Brewer (right) from the University of California, San Francisco...