... visualize their activity. In 2019, the first generation of norepinephrine sensors was developed, named...norepinephrine (Feng et al., 2019). But this first generation of tools had limited sensitivity and speed and... models of nLightG (left) and nLightR (right) generated using ColabFold (Mirdita et al., 2022). B, Representative...Speed One of the major advantages of this next generation of norepinephrine sensors is their speed. nLightG...GRABNE were more sensitive than nLightR, both generating a measurable signal after electrical stimulation...test point, ten pulses. This is in line with the general findings that red fluorescent proteins tend to ...improvements. In particular, they’re focused on generating variants with improved affinity and dynamic range...

...process, I’ve provided guidance and mentoring to several people that found my insights helpful. All went...of candidates either get no interviews, or get several interviews but no job offers, is a failure to clearly...understand your proposal to give you proper consideration. 3. Present an easy to follow seminar See item...techniques. You DO need to be prepared to know the literature of your field and all of the caveats and alternatives...human health. Failure to do so is implicated in several human diseases. Proper patterning of cells and ...that we can manipulate to study pathology and therapies." This is only the first few minutes of the...incorporated example documents. This part has generated a lot of positive feedback. Many of these documents...

...believed that the exonuclease activity of the polymerase generates sticky ends in the PCR reactions. These ...amplification of a recombinant DNA without sequence alteration. Molecular cloning techniques are some of the...its advantages, most scientists still encounter several struggles with these techniques: unwanted mutations... excessive PCR cycles or low fidelity Taq DNA polymerase, the construction of specific sequences for base...any subsequent steps. Using high fidelity DNA polymerases, I have never had any mutations introduced into...efficiency is highest when using high fidelity DNA polymerases like phusion. An 18-cycle PCR leads to the most...

...language is ... The general representation of data is... The figures are ... In general, I think that this...the study to be replicated or omission from the literature. I think that delivering constructive critique...studying plant development and abiotic stress tolerance, I have been peer-reviewing journal submissions... detailed. I prefer to do it on another day - preferably the following morning - as usually deadlines ...language in the first (and clunky) version of my general comments and adjust them if my opinion changed ..., increasing the impact of the work, and as a collateral benefit, increasing the impact of the specific...

...address this, several years ago we partnered with Seqwell to establish a next-generation sequencing (NGS... by Single-stranded Virus Next-generation Sequencing. Molecular Therapy - Nucleic Acids 4:e260 . https...of the VGS process and use cases in Human Gene Therapy. Existing methods for AAV quality control Historically...Viral Genomes in Academic Settings. Human Gene Therapy Methods 30:195–205 . https://doi.org/10.1089/hgtb...Consistency Evaluation of AAV Preparations. Molecular Therapy 25:1375–1386 . https://doi.org/10.1016/j.ymthe.2017.04.001...

...thoroughly answered by experts in human-computer interaction (HCI), and we have adhered rigorously to HCI ...qualities is an ongoing process that involves iterative refinements in response to customer feedback. ..., and then crafted a detection algorithm that tolerates occasional mismatches or indels. Empirical tests...effort to be rigorous, we dug into the original literature, some of it decades old, to provide reliable ...to the database. Another limitation is that by tolerating mismatches, our algorithm could annotate a feature...to newly sequenced strains through our mismatch tolerant detection algorithm and thereby speed up the annotation...the idea stage through all of its post-launch iterations.     Additional Resources on the Addgene Blog...

...gRNAs! Comic by Maya Kostman.   Important considerations before you start an experiment with CRISPR ...expression. Location and sequence are important considerations for designing your gRNAs. For indels, it's ...off targets. For CRISPRa and CRISPRi, these considerations are of roughly equal importance (target should...worry less about sequence optimality because you generally have fewer sequences to choose from). Finally,...non-homologous end joining (NHEJ) often leads to the generation of indels and thus frameshifts that disrupt the... the introduction of a specific mutation, one generally relies on homology directed repair (HDR) to incorporate...user may need to optimize numerous parameters to generate the desired edit. The wrench: Gene activation ...

...of proteins, and can be a readout for protein interaction studies such as immunoprecipitation and pull-... be assessed from a single western.  There are several variations of the protein transfer step including...wet, semi-dry, and dry but all follow the same general principle where the gel and membrane are “sandwiched... antibody should you use? You will likely find several different antibody options targeting your protein...example, in immunoprecipitation (IP) the antibody interacts with a protein in its native state while a western...are typically conjugated to an enzyme, such as horseradish peroxidase (HRP), that emits light upon reacting... cellular assays. They can determine protein interactions following immunoprecipitation and are a common...

...at once, virtual conferences may have apps or interactive calendars that you can use to help plan your ...highly voted questions,” says Zhang. “Usually the moderator starts by asking the speaker the highly upvoted...the Q&A in the chat box. Presenting? Make it interactive! As a presenter, it can be difficult to keep ...Workshop, Abitua and the other organizers found several avenues to keep the audience engaged. They first...Reproducibility for Everyone Workshop included this interactive activity during their session. Image from Reproducibility...Reproducibility for Everyone.  While this type of interaction might not work when you are giving a research...important to keep your video on and look into the camera to make it seem more like you’re connecting with...

...writing for a general audience. The tool highlights jargon terms or words rarely used in general writing in...university student.   The Pomodoro timer is literally a tomato! Image from Gerlos.   Tools for checking...desired formatting style. This tool is great for generating citations for a few papers at a time. Mendeley...version that works with Windows, Mac, and Linux operating systems.  Zotero  Zotero is also another free ...Can Grad School podcast episode 2 "Organizing Literature" : I found out about Zotero here Dear Grad Student...blog: Tips for writing scientific manuscripts: literature searching, reading, and organizing Tips for writing...