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Having demonstrated the practical utility of RNA-Amp™ using single cell inputs we next set out to further determine its characteristics by comparing the transcriptional profiles generated from RNA-Amp™ single cell samples to RNA-Amp™ cDNA from purified RNA equivalent to approximately 100 cells equivalent and unamplified reference RNA.
In each qPCR reaction, 100ng RNA equivalent were used.
The amount of MIR168a was 3.2 × 10−6 fmol (1 920 copies) per 100 pg of total RNA, equivalent to 853 copies per cell.
As dietary N increased, rumen bacterial purine bases [ribonucleic acid (RNA) equivalent] decreased linearly (P < 0.05), whereas total N content increased (P < 0.05).
Using agarose gel electrophoresis, the RT-LAMP assay detected as little as 0.1 fg of viral RNA (equivalent to 50 viral particle).
Modern cells instead have a protein-based enzyme called RNA polymerase (RNAP) that copies strands of DNA into their RNA equivalent.
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Three independent RNA extractions were carried out on this vector and RNA equivalents were determined by RT-qPCR.
An independent evaluation [10] of Transplex observed good comparability of differential expression to non-amplified RNA when Transplex used RNA equivalents of 1,000 or more cells (12 to 300 ng total RNA).
RNA equivalents were normalized to simultaneously determined glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA levels in each sample.
20 ng total RNA equivalents of cDNA were used in each RT-qPCR amplification run in triplicate.
RNA equivalents (RNA/ml), transducing units (TU/ml) and p24 concentration (pg p24/ml) were determined after concentration of LV by low-speed centrifugation.
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