...change their excitation/emission behavior upon environmental changes (e.g. pH, Ca2+ flux, etc). The most ...Oligomerization: The first generations of FPs were prone to oligomerize. This may affect the biological function... these FPs cannot be used in oxygen deprived environment. Recently, a new GFP isolated from the Unagi ...you are planning to express the FP in acidic environments (e.g. yeast cytosol, which is slightly acidic...

...DNA dot blot...Transduction assay...SDS-PAGE...Electron microscopy… Any guesses? That's right... ways...genome-containing viral capsids is typically quantified by electron microscopy of a viral vector solution. Since this... this technique is intensive and requires an electron microscope, it is not routinely performed on all...genome-containing (Figure 1).    Figure 1: Electron micrograph of an Addgene AAV viral vector preparation...

...contains iterons that serve as recognition sites for replication initiation proteins. These iterons vary ...plasmid with multiple oris that vary in DnaA-box and iteron structure, host-independent   Building broad...Development of constitutive and IPTG-inducible integron promoter-based expression systems for Escherichia...

...experimental systems. They demonstrated LOV-Turbo in neurons, in pulse-chase experiments, and more. Find LOV-Turbo...element (TRE) or the cFos promoter. For example, neurons treated with a phorbol ester activator gave fluorescent...Images of iPAK4 fibers in cultured rat hippocampal neurons. HaloTag dyes are switched from yellow to red at... Teaching an old antibody new tricks By Ashley Waldron While IHC protocols typically call for thin sections...

...Boyden (Addgene viral prep # 84445-AAVrg).   Electron Microscopy The ratio of empty to full (i.e., genome-containing...representative vector preparations was quantified with electron microscopy after negative staining. Empty vector...than full vector particles.     Figure 3: Electron micrograph of AAV vector preparation shows that...psPAX2 and pMD2.G packaging system developed by the Trono lab. Cell culture medium (OptiPro SFM) containing...

...refresher if you ever lose track of which acronym is which. Acronym Full name Notes   Base transition ...close at hand? Once again, this post has a lot of acronyms, so we've included a glossary at the end of this... for saturation mutagenesis, and they provide a strong proof of concept for further development. ...

...pluripotent stem cells (Chavez et al., 2015) into neuronal cells by activating genes that cause differentiation...CRISPR-based activation can be made inducible upon environmental triggers. This is done by creating a split dCas9...CRISPR activation are also reversible: once the environmental trigger leaves, gene expression is no longer...Dow D, Buser-Doepner C, Bertotti A, Trusolino L, Stronach EA, Saez-Rodriguez J, Yusa K, Garnett MJ (2019...

...either DTT or β-mercaptoethanol (BME). DTT is a stronger reducing agent than BME and has a less… distinctive...life when stored properly. Because BME is not as strong as DTT, you’ll have to use different concentrations...quantifying your blot. CCD cameras have a much higher upfront cost, but require no consumables, produce digital...like Coomassie Blue or Ponceau S. It is far less prone to error than normalizing to so-called housekeeping...happy blotting! Many thanks to Addgenies Ashley Waldron, Meghan Rego, and Amrita Rhodes for their help ...

... methods.   Rapid detection of 2019 novel coronavirus SARS-CoV-2 using a CRISPR based DETECTR assay...in the saliva (Lucia et al., 2020). Detecting coronavirus using CRISPR-based SHERLOCK (Specific ​H​igh ​...ultrasensitive and visual detection of novel coronavirus SARS-CoV-2 Sensitivity: 4.6-11 copies/microliter...2020). An ultrasensitive, rapid, and portable coronavirus SARS-CoV-2 sequence detection method based on...

...reports 5 (2015). BIOFAB plasmid set 3000 Series Confronted with the problem of creating vectors for predictable...Shine-Delgarno sequences used in this so-called bicistronic design (BCD). The 49 vectors in this kit can ...of varying strengths centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013...terminators can be used to precisely tune your polycistronic E. coli expression constructs. Some applications...

...aggregation in your prep. If your proteins are the wrong size on your gel or blot, Table 1 (at the end of...and keep an eye on it and the leading line (dye front) visible in the sample buffer. You’ll want to run...interest, but you don’t want to run so long your dye front or ladder runs off the gel. Luckily, this is easily...easily monitored in real time, as the dye front (and many ladders!) are visible to the naked eye.   If ... Troubleshooting Suggestion Proteins are the wrong size Proteins may be aggregating Lyse at lower...

...to reduce false positives within a test.   The Bonferroni Correction and the Benjamini-Hochberg procedure...control for large numbers of comparisons. The Bonferroni does this by establishing a significance threshold...However, if many comparisons need to be performed, Bonferroni and Benjamini will greatly reduce family-wise...Dependent Variables: A Monte Carlo Simulation Study. Front. Appl. Math. Stat. Volume 2 (2016). doi.org/10.3389...

...many scientists operate in a hyper-competitive environment where getting hired hinges on performing near...government scientists to the de-funding of crucial environmental research programs, Canada has been here before...researcher in the University of Michigan Department of Astronomy. She studies how planets form around young stars...community of science supporters is much larger and stronger that I expected. Jennifer Stancill, Ph.D., is ...

... Studying cell mobility within these complex environments has never been easy and, for many years, researchers...same laser) cooperate to cause a higher-energy electronic transition in a fluorescent molecule (see figure...adaptive and innate immunity in diverse tissue environments. These studies provide quantitative measurements...-parasite interactions in the brains of mice chronically infected with Toxoplasma gondii." The Journal...

...Heinrichs, 2009). When a fluorophore absorbs light, electrons are elevated from a ground state to an excited...After a brief elevation in this excited state, the electron returns to its ground state by emitting a photon...Pyrgiotakis, G., Zhang, Z., Purington, E., Gramlich, W., Grondin, Y., Rogers, R., Bousfield, D., & Demokritou, P...nanocellulose for nanotoxicological studies. Environmental Science: Nano, 6(5), 1516–1526. https://doi....

... (3): An optics system A fluidics system An electronics system  Cells are first resuspended in a pressurized...fluorophore or fluorochrome-stained protein will emit a stronger photon than a cell expressing low levels of the...triton-x, saponin, or tween. Take care when using strong detergents like triton-x and tween as extended ...FACS In addition to the fluidics, optics, and electronics systems, FACS instruments have specialized components...

...in E. coli by co-expressing the molecule chaperone Skp. Front Cell Infect Microbiol 3: . https://doi.org...Antibodies: Historical Perspective and Future Outlook. Front Immunol 8: . https://doi.org/10.3389/fimmu.2017.01589...biosensor biorecognition elements. Biosensors and Bioelectronics 85:32–45 . https://doi.org/10.1016/j.bios.2016.04.091...

... present in the sample. Other techniques, like electron microscopy and FRET, can offer more information...well described in the closed conformation, recent electron microscopy studies have also found Cas9:RNA complexes...pairing in the seed sequence.   Figure 6: Electron microscopy structure of S. pyogenes Cas9 bound...structure (left, PDB ID 4ZT0) with Cas9:RNA:DNA electron microscopy structure (right, PDB ID 7S4X), viewed...

...particularly energy-intensive systems in a more environmentally responsible manner.  Other ways to make the...in-depth look at Addgene’s current lab-oriented environmental efforts check out Leila Haery’s blog series,... we're always striving to set an example of environmental stewardship.  Additional resources on the Addgene...

... connections between BLA projection neurons and mPFC interneurons can be labelled with an AAV1 Cre virus...AAV9 tdTomato virus in the mPFC.  The AAV9 is so strong that you can even see dendritic spines!  Curious...