Step 5 - Programming The Plate Document Method - Applied Biosystems 7900HT User Manual

Step 5 – Programming the Plate Document Method
User Access
Requirement
About SDS
Methods
Database
Considerations
Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
If using the SDS Enterprise Database, you must belong to the Scientist or
Administrator User Group to modify the method of a plate document.
During a run, the SDS software controls the instrument based on the instructions
encoded in the method of the plate document. Each new plate document
(except allelic discrimination) contains a default method that you must customize for
your experiment.
Methods contain the:
• Thermal Cycler Conditions
• Auto Increment Values
• Ramp Rates
• Data Collection Options
• Reaction Volume Setting
To create a method for...
• absolute quantification
• relative quantification
allelic discrimination
dissociation curve analysis
Configure the method of your plate document to collect data only in the necessary
stages of the PCR. Because of memory constraints, the SDS Enterprise Database
cannot save real-time plate documents greater than 40 MB. The file size for a
real-time run depends on the length of the run and the configuration and placement
of the data collection icons. Longer runs and runs configured to collect data at
multiple stages of the method can be considerably larger than the 15–25 MB average.
Step 5 – Programming the Plate Document Method
Then...
program the method as explained on
IMPORTANT!
If using the SDS Enterprise Database, read
the discussion of considerations below.
see page 3-20.
To perform a dissociation curve analysis do the following:
1. Program the method for the absolute quantification
experiment as explained on
2. Add a temperature ramp to the method for dissociation
curve analysis as explained on
DRAFT
September 1, 2004 11:39 am, CH_Run.fm
page 3-21.
page 3-21.
page 3-23.
3-19