The instrument requires valid ROI/uniformity, background, and dye calibrations to perform instrument
verification.
Instrument verification description
Purpose
Description
Pass criteria
Confirms the
performance of the
instrument.
Quantifies the number of copies of
the human RNase P gene in samples
with known concentrations of the
corresponding DNA template.
The instrument passes performance specifications
if the following inequality is true and the
instrument successfully distinguishes between
unknown populations A and B with a statistical
confidence level of 99.7%.
[(C
tA
) – 3(σ
CtA
)] > [(C
tB
) + 3(σ
CtB
)]
where:
• C
tA
= Average C
t
of unknown population A
• σ
CtA
= Standard deviation of unknown
population A
• C
tB
= Average C
t
of unknown population B
• σ
CtB
= Standard deviation of unknown
population B
RNase P instrument verification plate
The RNase P plate contains the reagents necessary for the detection and quantitation of genomic
copies of the human RNase P gene (a single-copy gene encoding the RNase moiety of the RNase P
enzyme). Each well contains: PCR master mix, RNase P primers, FAM
™
dye-labeled probe, and a
known concentration of human genomic DNA template.
1
4
2
5
7
6
3
8
Figure 1 96-well RNase P plate
1
Unknown A (5,000)
2
NTC (no template control)
3
STD 1,250 copies
4
STD 2,500 copies
5
STD 5,000 copies
6
STD 10,000 copies
7
STD 20,000 copies
8
Unknown B (10,000)
Chapter 5
Calibrate and verify instrument performance
Perform instrument verification using RNase P plates
5
QuantStudio
™
1 Plus Real-Time PCR System Installation, Use, and Maintenance Guide
61
