Skip to main content

Table 2 Template comparison

From: A novel, non-radioactive eukaryotic in vitro transcription assay for sensitive quantification of RNA polymerase II activity

Template qPCR C(T) values qPCR C(T) values
No RNA purification RNA purification
Positive Negative Positive Negative
CMV-EGFP Free 32.6 31.6 17.4 >40
Beads 37.3 >40 23.2 >40
HS-DNA Free 27.7 22.3 30.3 37.0
Beads 28.1 29.6 20.7 >40
  1. Two different linear DNA templates, CMV-EGFP or HS-DNA in solution (free) or immobilized on magnetic beads (beads) were used. Only positive transcription (+) and negative control reactions (−) were performed, and the product detected using primer extension and qPCR. C(T) values for the positive reactions should be lower than 25, values over 30 for the positive reactions reflect RNA synthesis levels that are too low to allow reliable inhibition quantification. Negative reactions should result in C(T) values higher than 35. C(T) values > 40 reflect no amplification, i.e. no detectable RNA synthesis.