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..., we've looked back on the year would like to highlight some of the top articles from the Addgene blog...-CoV-2 protein expression, and more. We also highlighted reagent repositories and other collaborative ...time so that you can begin your cloning experiment right away. And without further ado, here are the top ...about hosting multiple plasmids in one cell, it might be that you can't stably maintain them in the cell...free molecular biology tools out there! Here, we highlight a subset of these tools for plasmid mapping, DNA...

...purposes Rights to all research results, modifications, and inventions created by the Recipient Right to ...terms, so you can rest easy knowing your rights – and the rights of your institution – are protected while...scientist working hard on research, and you need the right experimental materials. Who has time for paperwork...materials. Questions about intellectual property rights, pending publications, and commercial use often...patent applications on modifications or inventions Right to publish without editorial comment or review by...order, Addgene generates the MTA and emails it straight to you, or we can send it directly to your institution...

...but some details might vary.  How it works — briefly Spectrophotometers measure how light travels through...through a sample — in this case, the absorbance of light at three key wavelengths: 230 nm, 260 nm, and 280...comparison. Each of these contaminants will have a slightly different impact on the spectra of a mixed sample...molecules that may be present in the sample also absorb light in this range of wavelengths (Figure 1B-D). These...remove from your sample is protein. Proteins absorb light to varying degrees across the UV range used in NanoDrop...This property means that even samples with quite high levels of phenol (up to 600 ppm according to Loughrey...difficult by spectrophotometry. RNA tends to yield a higher A260/280 and a lower A260/230 than DNA, so shifts...

...marched in solidarity with our American neighbours, the fight is far from over here at home.  The work...of which impact reproductive rights and justice. Whether it’s used to fight the myths that abortions cause...wishful thinking. The March for Science is about fighting scientific partisanship, including more scientists...there's been a disturbing trend in the number of high profile people denying science, from celebrities...convince me to spend a Saturday in the tourist-trap nightmare that is Times Square. But I made the sacrifice...Most people won't argue with that idea, but they might feel disconnected from the entire scientific process...I think many of us grew to assume that the data might speak for itself but we're now in a rift where our...

...collection of AAV enhancer plasmids with high specificity and brightness, now available at Addgene!   {% module_block...winners, containing enhancers with the highest specificity and brightness. You will find enhancers to target...allocated tremendous time and resources to generate highly specific tools to target different cell populations...border-style":"none","padding-left":"172px","padding-right":"172px"}{% endraw %}{% end_module_attribute %}{...font-family":"helvetica","font-style":"normal","font-weight":"normal","text-decoration":"none"},"fallback":...font-family: helvetica; font-style: normal; font-weight: normal; text-decoration: none","font_set":"DEFAULT...best-in-class collection. Hopefully, this collection of highly specific AAV enhancers can help scientists around...

...RNA molecules, which could bind their targets with high specificity and affinity, were eventually called...nucleotide-based ligand binders would prove to be highly adaptable tools. The key to aptamers’ flexibility...conformational changes after target binding, aptamers are highly customizable in terms of how they actually “report...applications. Learn About Fluorescent Biosensors Light-up aptamers (or, fluorescing with veggies) Though...eventually evolved into Spinach2, a more stable and brighter version of the aptamer, but it still had limitations...more stable in cells. The new aptamer also has a higher melting temperature than its Spinach ancestors,...have also made aptamers an important component of high throughput assays with microarrays and biochips....

... client situations might be especially challenging. For instance, the client might need to get a large...what a career in this field involves, and why you might enjoy doing it. I’ll also talk about the skills ...and anticipated commercial success. Or the client might need help with the intricate task of designing a...topics are typically cutting edge; a consultant might outline the impacts of combining targeted molecular...turnover of projects, pressure to maintain consistently high levels of performance, and occasionally being faced...cases to better staff future teams. As with any highly collaborative work place, challenges can arise ...or medical device works. This understanding sits right at the core of the suite of benefits a given therapeutic...

...fluorescent biosensor has a high sensitivity (50 nM-500nM range) and a high specificity for SLs in vitro...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...organoids. The Organoid EasyTag System is a straightforward workflow for doing gene targeting, reporter.... This novel type of biosensors could be used in high throughput screening for agrochemical compounds ...CRISPR Plasmids and Resources page. Here are some highlights from the past couple months: A new lentiviral...

...activation (chemogenetics) or light (optogenetics).   Timing Optogenetics offers a high degree of temporal resolution...presence or absence of the correct wavelength of light. This response is immediate, on the scale of milliseconds...tissue-subregion targeting down to the limits of light delivery. Chemogenetic approaches excel at broad...offers better control over these factors as the light source can be easily and quickly manipulated or ...Addgene website. Optogenetics also requires that light of the correct wavelength reach receptors in the... address this is by working with organisms with light accessible brains, such as zebrafish larvae. Mouse...implant to be surgically installed to allow for light delivery. Chemogenetics poses the advantage of being...

... the high school interns as equals, because as soon as I had passed my training, I dove straight into ... never would have imagined that my summer before high school senior year would have been spent within ...weeks that we had made a mistake by not having the right concentration of a reagent during our HiFi assembly...the following two weeks, we were able to obtain eight clones. We ran digested and undigested samples of...one clone we found with the correct bands was the right plasmid, so after discussion, we sent it for sequencing...invited guests came to see the research from all the high school interns, done within the past six weeks. ...other way. Toj Toj-Villagran is a rising senior in high school who just completed their summer internship...

... tips for high efficiency cloning with this technique: Amplification efficiency is highest when using...labs, a variety of other cloning techniques with higher efficiency and simpler procedures have been developed... in the figure from Chaokun et al., 2011 to the right. After PCR, you should confirm that the products...introduced during PCR or any subsequent steps. Using high fidelity DNA polymerases, I have never had any mutations...chance of introducing an unwanted mutation. Tips for high efficiency cloning Now that you have a better understanding...using high fidelity DNA polymerases like phusion. An 18-cycle PCR leads to the most colonies after transformation...27391134. 2. Li, Chaokun, et al. "FastCloning: a highly simplified, purification-free, sequence-and ligation-independent...

...neurons in the mouse brain. MouseLight images can be found on the MouseLight website. Together with scientists...neuroanatomy and neural connectivity, providing valuable insights for neuronal communication. Viral vectors and...vectors can be equipped with strong promoters for high and neuron-specific transgene expression. Fluorescent...for their production and purification that yield high titers (see also AAV protocols for production, purification...researchers from the Janelia Campus for their MouseLight project but neurons were traced with AAV2. Two-Photon...variant, rAAV2-retro, that migrates retrogradely at higher rates than any other AAV serotype (Tervo et al....also having the added advantage of lower toxicity, higher specificity and reduced technical variability compared...

...forward, in this post I’ll point out some of the highlights and takeaways I got from the symposium. The ...outcome is not easily achieved. Of note, it was highlighted during the conference that just increasing funding...projects sought to make an interactive game that highlights just this point. As part of this, trainees need...thoughts The FOR Symposium Boston 2015 shined a light on some of the many problems facing the academic...academic research enterprise. In addition to highlighting some of the important questions that still need to...

...Some common ones you might see include ColE1, pMB1 (which comes in a few slightly different but well known...make additional copies. ori sequences are generally high in As and Ts. Why, you ask? Well, A-T base pairs...make a lot of your plasmid? Is the gene toxic in high amounts? It is always good to keep in mind that ...propagate plasmids, but endA- E. coli are best for high yields of plasmids. Growth conditions The amount...culture inoculum Freshly streaked bacteria have higher copy numbers – for optimal results always pick ...cultures. Freshly transformed cells also give a higher plasmid yield that colonies streaked from a glycerol...stock. Incubation for 12-16 hours tends to give higher copy numbers since the bacteria have just reached...

...with different properties (for example, overall brightness or on/off kinetics) that makes them suitable ...(pGP-AAV-syn-FLEX-jGCaMP7b-WPRE) exhibits the brightest resting fluorescence of the GCaMP7 family of calcium...pAAV.hSyn-FLEX.iGABASnFR) has good membrane localization and brightness. The F102G variant (pAAV.hSyn-FLEX.iGABASnFR....through imaging. Since it’s illuminated by red/orange light, Archon1 can also be used in conjunction with optogenetic... activators and inhibitors that respond to blue light. We recently released a constitutive (pAAV-Syn-Archon1...-jGCaMP7f-WPRE AAV1 with a CAG promoter exhibits high specificity for Purkinje neurons. 105321  AAV1...pGP-AAV-syn-jGCaMP7c variant 1513-WPRE 7c exhibits high contrast between peak fluorescence and resting fluorescence...

...While I had always enjoyed learning about biology in high school, it wasn’t until I started working in my ...molecular biology-based activities making their way into high school classrooms, like gel electrophoresis to separate...led me to develop BioBits ExplorerTM and BioBits BrightTM, biology kits based on freeze-dried, cell-free... that students can easily sense, whether that’s bright fluorescent proteins that they can see, enzymes...initially developed the BioBits kit to be used in high school classrooms, but it could be widely adapted... a biology teacher at the elementary, middle, or high school level, feel free to check out one of our ...PMC6070312. 2. Stark, Jessica C., et al. "BioBits™ Bright: A fluorescent synthetic biology education kit....

...projects in many labs. However, the high costs of sample preparation for high-throughput next generation (NGS...Addgene to develop a highly optimized bioinformatics pipeline that will return high-quality assembled and...quantities of library molecules for each sample remain highly normalized across a wide range of input DNA concentrations... vary by DNA input amount. plexWell™ + Addgene = high quality NGS-assembled plasmids Because of the enhanced... plasmids. The plexWell™ workflow also offers a highly efficient NGS-based alternative to Sanger sequencing...assembly pipeline. To date, this has resulted in high quality NGS-based assemblies for over 3000 plasmids...

...fluorescence changes or a requirement for very high excitation light intensities have made these indicators difficult...Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids...measure ApoB-LPs directly and develop a potentially high throughput in vivo model to identify drug candidates...Thierer et al, 2019). Zebrafish are a great model for high throughput study of ApoB-LPs because their ApoB ...amenable to use with other bEBP/UAS pairs, and displays high dynamic range. However, the promoter in question...lipid compositions of C. elegans membranes, which might beneficially influence the rhodopsin properties....

...commonly used, PVDF tends to be slightly more popular due to its durability and high binding capacity. In addition... (HRP), that emits light upon reacting with a specific substrate. The emitted light, or chemiluminescence... samples, a protein standard of known molecular weight in a separate lane can help confirm the size of...acrylamide content of the gel. For small proteins, use a higher percentage of acrylamide to increase migration ...broad range of protein sizes but takes hours to overnight.  In contrast, a semi-dry transfer only requires...chain. Consequently, antibodies validated for IP might not work for a western and vice versa.  In addition...specific experimental condition you could look for a slight size shift in the protein band before and after...

... likely has the blue-light photoswitching issue). Although similar in brightness to jRGECO1a and FRCaMP...Every few months we highlight some of the new plasmids, antibodies, viral preps, and more in the repository...biomolecular condensates AAV-CRISPRa-engineered fat cells fight cancer Frugal CRISPR kit for classrooms Cell-surface... to RCaMP3 and jRGECO1a, PinkyCaMP had better brightness, signal-to-noise ratio, photostability, and signal...mScarlet-based calcium sensor with exceptional brightness, photostability, and multiplexing capabilities...collaborators have found a way to make adipocytes fight cancer. In this method, called adipose manipulation...separately. Results can be analyzed using either "high-tech" or "low-tech" equipment. Reproduced from Collins...