...Deisseroth 83895 AAVrg pAAV-hDlx-Flex-GFP-Fishell_6 Gordon Fishell   Optogenetics Optogenetics continues...Cepko 83898 AAVrg pAAV-mDlx-ChR2-mCherry-Fishell-3 Gordon Fishell   Chemogenetics Chemogenetics offers...pAAV-hDlx-GiDREADD-dTomato-Fishell-5 83897 AAVrg pAAV-hDlx-GqDREADD-dTomato-Fishell-4 Controls Plasmid...pAAV-mDlx-GFP-Fishell-1 Biosensors Plasmid Serotype Name 83899 AAVrg pAAV-mDlx-GCaMP6f-Fishell-2 ...

...and an ever increasing number of journals and publishers to choose from, it gets very confusing for scientists...research may stipulate that your article must be published in an open access journal or by using the open...your manuscript will be available to anyone who wishes to read it, without them needing a subscription...data resulting from their funding should also be published. All these factors will influence your choice ...manuscript needs to be revised before it can be published, this can delay publication and manuscripts may...referee report are permanently attached to the published article. In addition, authors and readers are ...help your chances of getting your manuscripts published in the future. Disregard reviewer’s comments without...

...of the number of papers with your search term published each year, which is a nice way to see growth or...receive a daily, weekly, or monthly digest of newly published papers that contain that term. You can also set... a collection, which is essentially a virtual bookshelf. Pro tip The “cited by” and references sections...papers related to a particular topic or that were published by the lab down the hall.  Scientific journals...graph of the number of papers with your key term published each year is displayed with your search results...searches to particular authors, with the authors who published the most papers with your keyword populated at...Meta. Feeds automatically update as papers are published, similar to Facebook or Instagram. Key features...

...relevant to the phenotype being studied. Examples of published selection mechanisms for lentiviral CRISPR libraries...

...like ReFigure which aims to consolidate published and unpublished experiments on similar subjects in a single...the abbreviated methods section found within a published manuscript. Luckily, many organizations have been...thereby enabling you to do modified analyses on published data if you need to. Similarly, FigShare also ...averse to producing papers that repeat previously published results. This over-reliance on the perfect story...problem is that only these positive results will be published. The publication record will then show that the...success story) even though the majority of the unpublished evidence shows otherwise. Others will go on to...very active with many scientists sharing their unpublished data. There are blogs - both hosted by individuals...

...validation assays. If you’re using previously published antibodies, check the manuscript and supplementary...mitochondria or nucleus, will likely require a harsher lysis buffer than proteins easily available in ...in the cytoplasm or whole-cell lysate. A harsher, SDS-containing lysis buffer, like RIPA, should be considered.... BME smells like rotting eggs but has a longer shelf life when stored properly. Because BME is not as... 1–2 hours Easy and cheap to handcast Short shelf-life; can alter proteins due to high pH Bis-...for proteins <40 kDa 180 V, 30 minutes Good shelf life; runs at neutral pH (reduced protein alteration...for proteins > 30 kDA 200 V, 30 minutes Good shelf life; runs at neutral pH (reduced protein alteration...

...the transfection in: 15 cm dishes OR CF5 stacks Select how many dishes or stacks you plan to transfect...for the type of tissue culture dish and number of dishes used. Here’s to making AAV packaging a little easier...1 to 10 Fill in experimental details on the spreadsheet: Name your viral prep Enter the specific names... plasmids Enter plasmid concentration The spreadsheet will calculate the following μg of each plasmid...with its corresponding column in the calculator spreadsheet: We hope that this calculator will make it easier...

...Originally published May 3, 2017 and last updated Sep 24, 2020 This post was contributed by guest blogger...Gene knockout with CRISPR technology is usually accomplished by Cas9-mediated dsDNA breaks: following a cut...mRNA cap, provides the most accurate mapping (Radzisheuskaya et al., 2016). In this case, location and sequence...nevertheless still lead to activity has been established, and can be used to identify sites that are likely...Zhivich V, Najgebauer H, Allen F, Gonçalves E, Shepherd R, Doench JG, Yusa K, Vazquez F, Parts L, Boehm...Boehm JS, Golub TR, Hahn WC, Root DE, Garnett MJ, Tsherniak A, Iorio F (2019) Agreement between two large .... https://doi.org/10.1038/s41587-020-0490-7 Radzisheuskaya A, Shlyueva D, Müller I, Helin K (2016) Optimizing...

... but if available, use an automated ELISA plate washer. Equipment Spectrophotometer compatible with 96...controller Microcentrifuge Microplate shaker ELISA plate washer (optional) Scale Microsoft Excel or similar software... but if available, use an automated ELISA plate washer. Seal the plate. Place the plate on a microplate... . Repeat steps 8–10 twice for a total of three washes. After the final wash, aspirate the wash buffer... but if available, use an automated ELISA plate washer. Seal the plate Place the plate on a microplate.... Repeat steps 18–20 twice for a total of three washes. After the final wash, aspirate the wash buffer...

...pre-clinical life science work, economics) that published research is difficult to replicate.  Why is replicating...appreciate how few details are provided in most published papers. The precise timing, order, or every combination...to a preregistration, but usually with a fully fleshed out introduction section) to one of 222 participating...scientific research. These include policies for publishers and funders in the Transparency and Openness ...citizen science and behavioral ecology of cichlid fishes. Find David online at https://orcid.org/0000-0002...

...Originally published Sep 30, 2014 and last updated Dec 10, 2020 by Benoit Giquel.   Adenoviral vectors...oncogenicity or genotoxicity seen with other vectors (Lukashev et al., 2016). The protein capsid encapsulating...Manuel Gonçalves' lab found in a second paper published in Nature Methods the same year that delivering...1051–1057 . https://doi.org/10.1038/nmeth.3075 Lukashev AN, Zamyatnin AA Jr (2016) Viral vectors for gene...doi.org/10.1038/srep05105 Stephens CJ, Lauron EJ, Kashentseva E, Lu ZH, Yokoyama WM, Curiel DT (2019) Long-...

...eukaryotic cells as an intermediate of heme metabolism (Shemetov et al, 2017). Modern state-of-the-art NIR FPs ...Shcherbakova et al., 2018) and iGECI calcium indicator (Shemetov et al., 2021). In addition to FRET, intensiometric...daria_gift_shch.     References and resources References Shemetov, A.A., Oliinyk, O.S. & Verkhusha, V.V. How to ...scales. Nat Commun 11, 239 (2020). Oliinyk, O.S., Shemetov, A.A., Pletnev, S., Shcherbakova, D.M. & Verkhusha...near-infrared FRET. Nat Chem Biol 14, 591-600 (2018). Shemetov, A.A. et al. A near-infrared genetically encoded...

...for development into chemogenetic tools. Need a refresher on G-protein coupled receptors? Read the GPCRs...

... substantial overlap in the activation curves (Marshel et al., 2019) for each tool, and you may have to...the right information When using a previously published system in your own work, you’ll need to know (... Tye, K. M., Ferenczi, E. A., Ramakrishnan, C., O’Shea, D. J., Prakash, R., Gunaydin, L. A., Hyun, M.,...13(3), 387–392. https://doi.org/10.1038/nn.2495 Marshel, J. H., Kim, Y. S., Machado, T. A., Quirin, S.,...

... to replicate at a higher frequency (Summers & Sherratt, 1984; Summers et al., 1993). These larger plasmids...the monomeric version of the plasmid (Summers & Sherratt, 1984; Summers et al., 1993; Summers, 1998). Multimeric...of plasmid multimers (Bedbrook & Ausubel, 1976; Fishel et al., 1981). E. coli strains such as DH5α, HB101...10.1128/microbiolspec.PLAS-0025-2014. PMID: 26104344. Fishel RA, James AA, Kolodner R. recA-independent general...2958.1993.tb01648.x. PMID: 8361350. Summers DK, Sherratt DJ. Multimerization of high copy number plasmids...

...with your blocking, first check your antibody spec sheet. Some antibodies work better with specific blockers...include recommended blocking conditions on the spec sheet. If you’re still seeing high background after following...longer incubations at both temperatures if needed. Washes Check your antibody manufacturer for wash recommendations...consider adding a detergent like Tween-20 to your washes — although this can be harsh and inhibit some detection... Inefficient blocking Check antibody spec sheet for buffer recommendations Use a chemical blocking...signaling after each round of stripping* Use a harsher stripping buffer Too little signal Large ...

...yearly planning meetings revolve around two simple worksheets, each of which is filled out separately by the...the trainee and PI before the meeting. These worksheets, summarized briefly below (and available of in...facilitate honest conversation. Goals and Planning Worksheet: Lists accomplishments, research/professional...often give ourselves credit for what we have accomplished. Starting with a recap of accomplishments, from...

...CRISPR-based phenotypic screen datasets. Original published CRISPR screen data is curated and deposited in...in BioGRID ORCS to allow facile queries of published data by biomedical researchers and ready access to... approach serves to maintain consistency with published screen interpretations and allows users to query...curators at [email protected] to submit their published data for rapid curation review and upload. In ...

...wanted to do when I grew up until I was all but finished my undergraduate degree (and I hope I’m not alone...towards a career in science communication, an established field of inquiry with which I was not yet familiar... a scientific background and bring with them established interests and areas of expertise. Perhaps the...strongest aspect of science communication, then, is its sheer versatility in the face of disparate spheres of ...

...2013 there have been 33 CRISPR software tools published and documented by OMICtools. It must be confusing...The first of only three such databases, EENdb (published 1 Jan 2013, no longer available), is a simple ...sgRNA designs. The second database, CrisprGE (published 27 Jun 2015), is noteworthy for its broad scope... content, and ease of access. The third, WGE (published 15 Sep 2015), is more similar to EENdb than CrisprGE...in detail, but that is already covered in the published articles. Instead, I have broken down the tools...