...or the activity of a protein in a synapse. What kinds of tools enable us to measure these properties, ...articles describing the FRET tool that you are looking for and check our curated list of biosensors too... Sustarsic, Marko, and Kapanidis, Achillefs N. "Taking the ruler to the jungle: Single-molecule FRET for...

...alleninstitute.org. A classic challenge in cell biology is making sure that what we observe through the microscope...whole segment—a GFP tag and 1kb of homologous DNA flanking both sides (about 2.7kb total)—is inserted into...and destroyed by CRISPR/Cas9 during transfection. Making the double strand break and introducing the tag...

...Advancement of Science, Rush Holt, followed by a networking reception. Saturday’s workshop will be an advocacy...laid out by a 2014 National Academy of Sciences working group (National Academy of Sciences. “The Postdoctoral... Advancement of Science, Rush Holt, PhD and a networking reception. Saturday’s workshop will progress ...

... me troubleshoot some technical obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements...cells for infection If your transductions aren’t working, you can check if virus is being generated by your...template. Regardless of the type of virus you’re working with, there’s a lot that goes into virus design...

...recombination events, and that the orientation of the flanking loxP sites directs the Cre recombinase to invert...placement of the two pairs of target sites. By flanking the reporter with one loxP site and one lox511...PMID: 12665802. 2. Branda, CS and Dymecki, SM. Talking about a Revolution: The Impact of Site-Specific...

...density: particle concentration is measured by looking at absorbance at 260nm using UV/Vis spectrophotometry.... The purity of the sample can be estimated by looking at the ration of DNA (260nm) versus protein (280nm...calculated in terms of plaque forming units (pfu), taking initial vector dilution into account. You'...

...10.1038/s41587-023-01756-1. PMID: 37095348. Breaking up is hard to do: Monomeric StayGold variants ...with some applications. Several labs have been working to produce monomeric versions that break up the...was delivered to brain endothelial cells in mice lacking LY6A using AAV1-X1 (Chen et al., 2023); then, when...

...laboratory of Dr. Moshe Szyf at McGill University, working to develop CRISPR-based tools for epigenetic editing...Charlton J, Mattei AL et al (2018). Genome-wide tracking of dCas9-methyltransferase footprints. Nat Commun...Zhong C, Eason DE, Ross DL, Vachhani RV, Ming G-L, King JR, Song H, Sweatt JD (2013) TET1 Controls CNS 5...

...Scientists looking for more compact genome editors due to delivery constraints and scientists seeking a wider... prep in serotype AAV5! Get dLight1.3b here Looking for a knock-out in your experiments? AAV-flex-taCasp3...

...The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected...amount of virus you actually have in your prep. Taking time to properly titer your virus will not only...original plasmid which can over-inflate results. A packing vector deficient in reverse transcriptase will ...

... who keep changing their minds and have trouble making decisions. In my work as a thesis coach for graduate...categories. In addition, I also learned about two other kinds of difficult supervisors: Hands-off super busy ...explain how it is interfering with your work. Avoid talking about your emotions, such as frustration or anger...

...in brackets and add the upstream and downstream flanking sequences to their input string (Figure 2). This...prediction models. In addition, we provide options for nicking gRNAs (used for PE3 and PE5 prime editing strategies...strategies) based on a DeepSpCas9 score ranking (Kim et al., 2019). PRIDICT saves time and resources by ...

... I’m not going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs...the viral particles of the AAV serotype you’re working with are primarily found in the culture media, .... If you’re new to creating gradients, practice making the iodixanol layers before doing your first purification...

... localization, cell structure, intracellular trafficking, and protein turnover rates. Additionally, by...fission/fusion, nuclear import, and neuronal trafficking. Although FPs have enabled discovery of many ...cellular mechanisms, there are some limitations to working with FPs. Overexpression of fluorescently tagged...

...components of the indicator, which can make the kinetics of voltage sensing to signal output slightly slower...fluorescent signaler. The pro of this system is the kinetic response time from signal input to readable output...FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component counterpart and...

...the FP (fused to the protein of interest), thus tracking the localization of that biomolecule inside living...eel was shown to mature independently of oxygen, making suitble for use in anaerobic conditions. Maturation...fluorescent protein should I use?" And if you're looking for more fluorescence microscopy tools and techniques...

...our samples 1:6 million to 1:25 million.  After making the dilutions, they are transferred to a new plate...performing ddPCR Clean everything You must have a clean working area to titer your AAV by ddPCR. Droplet digital...generator, you should avoid pipetting too quickly when making your virus dilutions. This will reduce aerosols...

...domain proteins, they can be expressed in bacteria, making nanobodies a recombinant and renewable anti-IgG...available nanobodies from Pleiner et al. Before picking a secondary nanobody, it’s important to review ...expression plasmids here Additional Resources Looking for monoclonal primary antibodies to use with the...

...dye, mauvine, was discovered in 1856 by William Perkin. His discovery set off a flurry of activity and...labeling strategies and improved fluorophores are making chemical dyes increasingly attractive and accessible...Medical Institute. He would rather be in the lab making new fluorophores for advanced imaging modalities...

...does not do). Furthermore, molecular biologists working with vectors are, in most cases, interested in ... the viral integrase protein (IN) during step 3 (King et al., 2008) (Hu & Temin, 1990). At step 3, the....1016/0022-2836(73)90277-5. PubMed PMID: 4355596 King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C....