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In short, the technology combines horizontal gene transfer and expression with stringent screening technologies in a system where the same yeast cell is used to first generate the compounds and second to screen for activity of these compounds against an internal or external assay.
We found that VPS34-IN1 inhibited phosphorylation of PtdIns by recombinant insect cell expressed Vps34 Vps15 complex with an IC50 of ~25 nM employing both an in-house assay or using an external assay undertaken by Life Technologies Supplementary Figuree S1).
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Assays locally used by the blood bank for HBV and syphilis screening were run in parallel to quality-assured external assays supplied during the study, and all discordant samples were submitted to confirmation testing in reference laboratories in Mozambique and Belgium.
The need to increase the awareness for harmonization initiatives and external validation of assay performance is one of the general common findings of both initiatives.
Mouse liver gDNA was used as external standard to assay some of the common multiple house keeping genes like GAPDH, beta Actin and 18s rRNA in the samples and their geometrical mean was used to normalize the gene copy numbers and gene expression levels of the genes we assayed.
For the external pH shift assays, the conditions were the same as the H2 production assays except that the assay was conducted for 12 minutes, and KOH or HCl was added 6 minutes after the start of the assay to change the external pH from 5.5 to 6.5 or from 6.5 to 5.5.
For both assays, external quality control standards were within expected ranges.
The samples assayed for apoC-III in the external clinical laboratory were assayed in parallel for apoB-100 levels using immunoturbidity-based quantitative assay.
M. pneumoniae and C. pneumoniae quantified DNA controls (Advanced Biotechnologies Inc., Columbia, MD) were included in each assay as external positive controls.
Internal controls (stromal elements) were used to assess tissue integrity and external negative controls ensured assay quality.
When applied to quantitative real-time PCR assay with external controls, the DNA templates eluted from FTA Elute consistently led to lower cycle thresholds than DNA eluted from Whatman 903 for the same mock CSF dilutions (median reduction 2.2 Ct values, IQR 1.68) (Fig. 1).
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