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Monitoring extraction of these seven endogenous and three spike-in sequences extracted from 10 separate aliquots of the pooled plasma sample showed increased RNA yields from the QIAGEN RNeasy and miRNeasy kits compared to the other three protocols tested (Table 2).
As a result, the application of on-line NIR spectroscopy for monitoring extraction process in CHM could be very encouraging and reliable.
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Surrogate standards were added to each sample to monitor extraction performance and matrix effects.
An internal control is needed to monitor extraction efficiency and to detect inhibition.
The results of this particular application of implementing NIR spectroscopy to monitor extraction process are very encouraging.
The set-up is fully automated and features monitored extraction, which facilitates rapid optimization of the method.
An internal control consisting of phocine herpes virus was processed along with the specimens during the entire procedure and permitted to monitor extraction and amplification efficiency, including inhibition.
All samples were processed in the presence of internal standards to monitor extraction efficiency and verify measurement accuracy.
An internal standard of C3-caffeine (10 μg/ml) was added to the sample prior to extraction in order to monitor extraction and injection of each sample.
A human RNaseP gene PCR was used as an internal control to confirm the absence of PCR inhibition and to monitor extraction efficiency.
In GC MS experiments, to monitor extraction efficiency, 750 ng of internal standard controls (hexacosane (Sigma #241687) and triacontane (Sigma #263842) as dissolved in hexane) were added on to the glass plates immediately before pheromone extraction procedures.
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