...solutions are discussed below. There are numerous GFP variants. Here I use the name ‘GFP’, which refers to mEGFP...applications that require anaerobic conditions. In experiments requiring anaerobic conditions, proteins that...Cranfill et al., 2016 and FPbase.org). A number of variants remain fluorescent under acidic conditions, with...genetically encoded tag and exogenously added components. For a review see Lotze et al. (2016). To date...is used to excite GFP, may also excite several components that are naturally present in cells or media....although it still is worthwhile to generate several variants and check the functionality of the fusion (Mastop...

... can use AAVs to improve your genome editing experiments (with and without targeted nucleases). AAV genome...identified by PCR. Then, if necessary for subsequent experiments, the floxed neoR gene is removed via infection...researchers have devised methods to deliver CRISPR components as ribonucleoproteins (RNPs) that can be supplied...2017 recently showed that by delivering CRISPR components as RNPs and delivering two halves of a repair...possible sequentially repair a target site with the contents of the two AAVs. Using this technique, Bok and...can provide a powerful boost to genome editing experiments. We’re excited to see how you continue to use...

...have shown recently that desired genome-editing events by HDR can be generated more efficiently by (1)...as APP and PSEN1, in most cases the intended HDR events were corrupted by unwanted indels on the same allele...correctly edited alleles, when screening reasonable amounts of single cell clones. Furthermore, CRISPR/Cas9...controlled over-active Cas9 and repeated our experiments with blocking mutations, we still weren’t entirely...Alzheimer mutations we study are heterozygous in patients. As the probability of mutation incorporation ...the Addgene Blog Learn about High Fidelity Cas9 Variants How to Design Your gRNA Delivering CRISPR Ribonucleoproteins...

...for synaptic glutamate response. They generated variants optimized for different applications (v82 has ... and powerful high-throughput screen to select variants based on performance metrics specifically for ...Farahani PE, et al. pYtags enable spatiotemporal measurements of receptor tyrosine kinase signaling in living...BeadScan platform for screening candidate biosensor variants for performance under two-photon fluorescence ...temperature indicator B-gTEMP designed to meet those requirements (Lu et al. 2022). By fusing two bright fluorescent...about your favorite new biosensor plasmids in the comments below!   Additional resources on the Addgene ...

...and recombination events). TIDER can screen for specific homology-directed repair events and small edits...significant number of cells have been edited. For many experiments, the end goal is to generate a clonal cell line... Fig. 2: Size screening for large deletion events.   Validating knock-in mutations You’ve introduced...combo and are hoping for homology-directed repair events, or maybe you’re performing a single base pair ... flow sorting or drug selection, assuming your reagents are amenable to this. If delivery isn’t an issue...

... question you think rotation students should ask senior graduate students? 11:51 - 13:28 : How do you ..., we sit down to talk with two senior graduate students, Ben Vincent and Niroshi Senaratne, from the Biological...down of graduate student life. Senior graduate students can sometimes be elusive or have a certain mystique...program. Don’t be fooled! Most senior graduate students are friendly and full of useful advice. Stay tuned...

...funding for research, and also to encourage women, students, and underrepresented minorities to study STEM...them share their lab experiences, findings, and experiments as articles written for general audiences. So...scientists and doctors with their questions and comments.  Lastly, we are planning to launch an e-magazine...Instagram every Sunday. Undergraduate and graduate students are more than welcome in contributing to The Addictive...scientists. Keep your message to the main three points of your story. Interested in learning more? Visit...

... more transparent. Second, SITN helps graduate students develop their communication skills and engage ...to a layperson. They also enjoyed mingling with students and other attendees and getting the chance to .... The future of DayCon DayCon and other similar events show how meaningful science outreach can be, both...both for scientists and the public. For students, these opportunities to improve our communication and...efforts to engage the public - let us know in the comments section! More information about SITN: DayCon...

..., we heard over and over again that the legal documents were one of the biggest hurdles for scientific...documentation that allows you to conduct your experiments with confidence. Benefits of the UBMTA: Peace...Even with all those benefits, we know that legal documents can get tricky. Here’s how Addgene helps: For ...materials have been requested and by whom. For Recipients: Streamlined Processing: When you place an ...So go ahead and use Addgene plasmids in your experiments. You know that they’re properly covered for when...

...and is useful for direct imaging of events in cells and brain explants. This construct is available through...Nano-Lanterns including monitoring multiple cellular events such as the dynamics of subcellular structures ...NanoLuc® luciferase Many intracellular sensors of events like calcium release have been made using fluorescent...PMC4942401. 5. Susuki, K., et al. (2016) Five colour variants of bright luminescent protein for real-time multicolour... more about FRET Tips for Using FRET in Your Experiments Learn more about Fluorescent Protein Biosensors...

...charge). Larger fragments of linearized DNA migrate slower than smaller linearized fragments. You can separate...have a full plasmid ready to be used in your experiments.     Alternatively, this whole process can be...sell fast digest enzymes that can digest large amounts of DNA in as little as 10 minutes, but check with...You then add DNA ligase to covalently link the fragments together at the expense of ATP (see below, covalent...Find the Perfect Empty Vector for Your Cloning Experiments Browse Our Molecular Biology Resources ...

...communicators. In this chapter we learn about some of their events - science seminars, science cafes, and the SITN...discussed below). Chpater 3: SITN Popularity and Events - Amy discusses some of the programs SITN runs ...scientists from a variety of fields as well as students from the ed school, design school, and Kennedy...short-form blog posts help readers think through current events. Chapter 6: The SITN Editorial Process -Vini discusses...balances working in the lab and working with SITN. She points to collaboration and delegating as keys achieving...

...science stories Zombie ants Ophiocordyceps unilateralis is a fungus that infects ants and controls their ...their behavior. These ants become “zombies” and leave their foraging trail. They climb upwards until they ...temperature and humidity favors fungal growth. The ants bites down and locks their jaws into a leaf, and... two cells. At the same time, it sucks out its contents (hence the name Vampirovibrio). What’s left of...

... CRISPR/Cas9" from the Journal of Visualized Experiments (JOVE). The video publication by Stuart Orkin...produce intrachromosomal deletions based on the experiments of Kim and colleagues using zinc finger nucleases...The first is the study of non-coding regulatory elements. Regulatory DNA function is typically inferred...exploring the function of non-coding regulatory elements in their native chromatin context. An alternate...paired DSBs is inversions [3]. These rearranged segments may also be informative as to genomic function...

... in vivo experiments, and nLightG was successfully trialed in two-photon in vivo experiments. nLight indicators... work on further improvements. In particular, they’re focused on generating variants with improved affinity...allows for increased options for multiplexing in experiments, including ones that study alpha2-AR. Selectivity...

...scale: Look at the mEmerald variants first, followed by the color variants. The mEmerald plasmids offer...corresponding plasmid pages (e.g. see Supplemental Documents here). If you’re not sure where your protein of... to be helping the Davidson lab to share their reagents. "We are proud to be able to bring such an important...work of Michael Davidson and his colleagues." Acknowledgments Thank you to Michael Davidson, Michelle Baird...

...Worcester Polytechnic Institute. Though some WPI students chose to go the solo route for their Major Qualifying... the first place. The famous discoveries and experiments I’d been learning about for years were usually...year isolating pheromones from various C. elegans mutants by myself, I would have slowly gone crazy. As it...substantially and have plenty of ideas for further improvements. Types of teams There are, of course, many ...series of tasks with precisely defined goals and endpoints. “Project” is a very broad term since a project...too much work If you are one of those graduate students putting in 60-80 hours a week in the lab, guess...

... the method is powerful and offers immediate measurements in a live animal that can be correlated to behavior...fluoresces when the receptor is bound by DA. Two variants of the sensor, dLight1.1 and dLight1.2, exhibit...peaks of DA release.  Using dLight1 sensors in experiments The Tian lab used dLight1 both in brain slices...receptors, respectively). Using GRABDA sensors in experiments The Li lab used GRABDA in drosophila, zebrafish...here! Overall these sensors are powerful new developments in the detection and measurement of neurotransmitters...

...workflow combines thoughtfully designed plasmid components with Fluorescence Activated Cell Sorting (FACS... known to trigger seed germination of parasitic plants, this approach was also used to build and validate...Qi's lab, allows simultaneous gene activation in plants.  FnCas12a, unlike other Cas12a nucleases identified...in moderate thermophiles.  These evolved SpCas9 variants have improved activity on NAG PAMS and reduced...sensor available in AAV8! Expand your imaging experiments with the Cre-OFF/FLP-ON vector 137128-AAV8: Ef1a-Coff...

...digestion, fragments are released from nuclei by a short incubation at 37 °C. These short DNA fragments can ...differential access to transcriptional regulatory elements is controlled during development using the Drosophila... preparation and high-throughput sequencing. Improvements to the CUT&RUN method and using it in your own...Henikoff lab published a preprint paper outlining improvements to the initial CUT&RUN protocol, the most significant...