Thermo Scientific QuantStudio 1 - Rnase P Instrument Verification Plate; Performance Specifications Pass Criteria; Prepare an Rnase P 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
76
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)
After the run, the software calculates average copy number values and standard
deviation values. The instrument passes performance specications if the following
inequality is true and the instrument successfully distinguishes between unknown
populations A and B with a statistical condence 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
The software automatically adjusts the threshold and omits a dened number of wells
from the unknown populations to meet the performance specications. To view any
omied wells, open the EDS le for the verication in the desktop software.
Materials required for RNase P plate preparation
• RNase P instrument verication plate
• Centrifuge with plate adapter; buckets cleaned before use
• Powder-free gloves
• Safety glasses
RNase P
instrument
verification plate
Performance
specifications
pass criteria
Prepare an
RNase P plate
Chapter 5 Calibrate and verify instrument performance
Perform instrument verification using RNase P plates
5
58
QuantStudio
™
1 Real-Time PCR System Installation, Use, and Maintenance Guide