Applied Biosystems 7900HT User Manual page 104

Chapter 3 Preparing a Run
Programming
Methods for Allelic
Discrimination
3-20 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
September 1, 2004 11:39 am, CH_Run.fm
To maximize instrument throughput, the SDS software does not provide the option to
thermal cycle allelic discrimination plate documents. Because allelic discrimination
experiments are end-point runs that do not require data collection during the PCR,
you can perform thermal cycling on a dedicated thermal cycler and then transfer to
the 7900HT instrument for data collection and analysis.
If you want to thermal cycle the
allelic discrimination plates on...
a designated thermal cycler
the 7900HT instrument
Performing Thermal Cycling of Allelic Discrimination Plates on the 7900HT Instrument
To perform the thermal cycling and the plate read using the 7900HT instrument, run
the plate first as a real-time plate document and then again as an allelic
discrimination plate document as explained below.
IMPORTANT! Follow the procedure below only if you intend to perform the PCR on
the 7900HT instrument. Otherwise, perform the PCR on a dedicated thermal cycler
and then transfer the plate to the 7900HT instrument for data collection.
1. Start the SDS software.
2. Create a real-time plate document for absolute quantification as described on
page 3-9.
Note: It is not necessary to configure the plate document with detectors.
3. Program the plate document method with the thermal cycling times and
temperatures for your protocol as described in
Absolute or Relative Quantification" on page
4. Save the plate document as explained on
Do not save the real-time plate document to the database. The
IMPORTANT!
database cannot store plate documents of different run types with the same bar
code.
5. Run the plate using the real-time plate document as described on
Although large, the real-time file may be helpful in diagnosing and
Note:
troubleshooting the experiment later if the data from the allelic discrimination
run produces unexpected results.
6. When the run is complete, close the absolute quantification plate document.
7. Go on to "Step 6 – Saving the Plate Document as a Template" on page
DRAFT
Then...
go on to "Step 8 – Applying Sample and Plate
Information" on page 3-28
follow the procedure below.
"Programming Methods for
3-21.
page 4-24.
page 4-25.
3-24.