...like ReFigure which aims to consolidate published and unpublished experiments on similar subjects in a single...the abbreviated methods section found within a published manuscript.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 The publication record will then show that the...success story) even though the majority of the unpublished evidence shows otherwise. Others will go on to...very active with many scientists sharing their unpublished data.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...

...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...

... 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 ...