Applied Biosystems 7900HT User Manual page 266

Chapter 7 Maintaining the Instrument
Running the
Verification Plate
or TGF-β Card
Verifying
Instrument
Performance
7-34 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
September 1, 2004 11:39 am, CH_Maintenance.fm
1. In the plate document in the SDS software, select the Instrument tab.
2. In the lower portion of the Instrument tab, select the Real-Time tab.
3. If the instrument tray is inside the 7900HT instrument, click
The instrument tray rotates to the OUT position.
4. Place the TaqMan RNase P Instrument Verification Plate or TGF-β card into the
instrument tray.
The A1 position is located in the top-left corner of the instrument tray.
Note:
5. In the Real-Time tab, click
Before starting the PCR run, the instrument may pause (up to 15 min) to
Note:
heat the heated cover to the appropriate temperature.
6. When the run is complete:
a. Analyze the run data as explained on
b. Set the baseline and threshold values for the analyzed data as explained on
page 6-46.
c. Verify the performance of the 7900HT instrument as explained below.
Analyzing Data from RNase P Runs
The install specification of the Applied Biosystems 7900HT Fast Real-Time PCR
System demonstrates the ability to distinguish between 5,000 and 10,000 genome
equivalents with a 99.7% confidence level for a subsequent sample run in a single
well.
The following equation verifies the 7900HT instrument install specifications:
[ ( ) 3 σ (
CopyUnk –
1 CopyUnk1
where:
• CopyUnk is the Average Copy Number of Unknown #1 (10,000 replicate
1
population)
• σ
is the Standard Deviation of Unknown #1 (10,000 replicate population)
CopyUnk1
• CopyUnk is the Average Copy Number of Unknown #2 (5000 replicate
2
population)
• σ
is the Standard Deviation of Unknown #2 (5000 replicate population)
CopyUnk2
The values above can be obtained from the experimental report window.
Note:
Up to six wells from each replicate group in a 96-well TaqMan RNase P
Note:
Instrument Verification Plate (both standard 96-well and Fast 96-well plates) can be
ignored to meet specifications.
Up to 10 wells from each replicate group in a 384-well TaqMan RNase P
Note:
Instrument Verification Plate can be ignored to meet specifications.
DRAFT
. The 7900HT instrument begins the run.
page
) ] [ ( ) 3 σ (
> CopyUnk –
2 CopyUnk2
6-12.
) ]
.