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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.