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In terms of clinical application on the NASBA-ELISA, 200 μL of plasma samples (restored to room temperature from −80 °C) were used for RNA extraction by the same kit as above.
After RNA extraction, a DNA digestion was performed, and RNA was cleared by the same kit.
Cell lysates were collected as above and assayed by the same kit.
We extracted the same amount of the sequencing reads amplified by the same kit except the sequencing platform type to control the variables.
Activated caspase-3/7 levels were determined using the ApoTox-GloTM Triplex Assay (Promega) and normalized against the percentage of viable cells, also determined by the same kit, according manufacturer's instructions.
To evaluate the bias in the comparison caused by the correctable sequencing errors from Hiseq and Miseq, we extracted the same amount of the sequencing reads amplified by the same kit but sequenced on Hiseq 2000 or Miseq, respectively.
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For miRNA expression analysis, cDNA was synthesized with stem-looped miRNA-specific reverse transcription primers by using the same kit.
Viral RNA from sandflies was isolated by using the same kit, with minor modifications.
cDNA levels of B1R and B2R, as well as 18S rRNA were quantified by qPCR using the same kit and a DNA Engine Opticon 2 (BioRad).
The yeast cells were harvested at the OD600 = 1.0, and the total RNAs were extracted by using the MasterPure™ yeast RNA purification kit (EPICENTRE), and contaminated DNAs were removed by treatment of DNaseI in the same kit.
Data shown in Fig. 7b represent n=3 replicates (separate transfection experiments of the MODE-K cell line) measured in parallel to control for consistent Renilla and Luciferase ratios using the same kit; data were analysed by one-way ANOVA, with a Dunnett's post test, ****P<0.0001, ***P<0.001, **P<0.01, *P<0.05.
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