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This analysis was performed using the transcript levels and birth weights of the 48 individuals profiled on the whole transcriptome array.
However, these fold changes were calculated using the transcript levels at 5 weeks, i.e. when both strains reach sexual maturity, as baseline, whereas chronological age at the second time point differed between the strains, i.e. old age is reached at 14 weeks in GRZ but at 31 weeks in MZM-0403.
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Quantitative reverse transcription PCR (QRTPCR) was used to analyze the transcript levels of key genes involved in cell cycle processes (ftsZ, dnaA), photosynthesis (psbA, pcb, rbcL), the circadian clock (kaiC) as well as transcription (rpoD).
To validate the MPGR expression data, quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to measure the transcript levels of CrWRKY49, CrWRKY50, CrWRKY51, and CrWRKY52.
Prior to the advent of microarray and RNA seq, other strategies including subtractive hybridization were used to determine the transcript levels of multiple genes (Agarwal et al. 2014).
Quantitative RT-PCR was used to study the transcript levels of genes potentially related to the described phenomena between day 8 and 35.
The oligonucleotide primers, used for evaluating the transcript levels of ABA1, ABA2, ABI1, ABI2, ABI3, ABI4, ABI5, RD20A, RD29B, CBF1, CBF2, CBF3, COR15A, in the real-time RT-PCR experiments, were applied as Ding et al. [33].
The standard was used to obtain the transcript levels.
The comparative cycle threshold method was used to measure the transcript levels.
DNA microarrays are now routinely used to monitor the transcript levels of thousands of genes simultaneously.
The formula 2 REFERENCE CT TARGET CTCT) described in Chen and Dubcovsky was used to linearize the transcript levels for all genes.
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