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The neutral red retention (NR) assay was performed according to Borenfreund and Puerner[86] with slight modifications as detailed in Heger et al.[87] by using RTL-W1 cells.
The preparation of hippocampal cultures was performed essentially as described by Goslin et al. [48] with some modifications as detailed in Dresbach et al. [49].
This assay was performed according to the method of Borenfreund and Puerner (1985) with modifications as detailed: a 0.4%.
E. coli ribosomes were purified and assays were performed essentially as described by Tate and Caskey (1990) with modifications as detailed in reference Soleimanpour-Lichaei et al (2007).
The preparation of hippocampal cultures was performed essentially as described by Goslin and Banker (1991), with some modifications as detailed in Dresbach et al (2003).
Total genomic DNA was extracted from tissue samples of the foot preserved in 100% isopropanol following the protocol proposed by Sokolov [ 42] with slight modifications as detailed in Sauer & Hausdorf [ 22].
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Image analysis proceeded, with slight modification, as detailed elsewhere [55].
The assays were performed according to manufactures instructions with some modification as detailed in our previous work.
Similarly, we failed to confirm other positive reports of associations with hormonal risk factors, or of effect modification, as detailed below.
ChIP was performed essentially as described (Stock et al., 2007) with some modifications used for specific antibodies, as detailed in the Extended Experimental Procedures.
Although the work is definitely of eLife quality, the manuscript would benefit from some modifications, further experiments and reorganization, as detailed below: Major comments: 1) We are not sure if the microarray/ChIP-Seq analysis adds much to the paper.
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