Array card RNase P kits
The RNase P Kits include one empty array card and eight tubes of solution. Each tube
contains reaction mix (PCR master mix, RNase P primers, and FAM
™
-MGB dye-
labeled probe) and a known concentration of human genomic DNA template.
To perform an instrument verification
run, each solution is loaded into the
empty array card in the arrangement
shown in Figure 7. When complete, the
array card contains five replicate groups
of standards (200, 400, 800, 1600, and
3200 copies), two of unknown
populations (800 and 1600 copies), and
one that serves as a no template control
(NTC).
The software performs the same analysis of data from an instrument verification run
regardless of the type of consumable used.
After the run, the software:
1.
Generates a standard curve from the averaged threshold cycle (C
t
) values of the
replicate groups of standards.
2.
Calculates the concentration of the two unknown populations using the standard
curve.
3.
Calculates the following to assess the QuantStudio
™
6 or 7 Flex System
performance:
[(CopyUnk
2
) – 3(σ
CopyUnk2
)] > [(CopyUnk
1
) + 3(σ
CopyUnk1
)]
where:
• CopyUnk
1
= Average copy number of unknown population A
• σ
CopyUnk1
= Standard deviation of unknown population A
• CopyUnk
2
= Average copy number of unknown population B
• σ
CopyUnk2
= Standard deviation of unknown population B
Note:
Unknown population A refers to the 5,000-copy population and unknown
population B refers to the 10,000-copy population in the RNase P plate.
Figure 7
RNase P standards array card
layout
Analytical
performance
Chapter 2
Calibrate the instrument
Verify the instrument performance
2
QuantStudio
™
6 and 7 Flex Real-Time PCR Systems v1.6 Maintenance and Administration Guide
63
