ABI Prism® 7900HT Sequence Detection System ... - OpenWetWare

Setup Checklists
Experiments/Runs
Performedonthe
7900HT Instrument
Absolute
Quantification
Checklist
See the appropriate page for the type of experiment or run you want to perform:
Experiment/Run See Page
Absolute Quantification 4-3
Allelic Discrimination 4-4
Background 7-13
Dissociation Curve (Melting Curve) 4-4
Pure Dye (Spectral Calibration) 7-17
RNase P Instrument Performance Verification 7-24
To create and set up a plate document for an absolute quantification run:
Done Step Description See Page
1 Create an absolute quantification plate document. 4-6
2a a. Create detectors for the absolute quantification probes. 4-7
b. Copy the detectors to the plate document. 4-8
3a a. Configure the plate document with detector tasks
(NTC, Standard, and Unknown).
4-11
b. Assign quantities to the wells of the plate document that
contain standards.
4-12
4 a. Program the method for the absolute quantification run. 4-13
b. If performing an assay in which you would like to collect
dissociation data, add a temperature ramp to the thermal
profiletoperformadissociationcurveanalysis.
4-16
5 Choose from the following:
If running… Then…
a single plate continue to step 7.
the first plate in a series Save the plate document as an ABI PRISM
of plates with identical SDS Template Document as explained on
assay configurations page 4-17.
6 Create a plate document from the template created in
step 5.
4-18
7 Configure the document with sample names and plate
information.
4-19
8 Prepare and run the absolute quantification plate or plates. 4-20
9 Analyze the run data. 6-3
a. Steps 2 and 3 can be eliminated by importing the plate document setup information from a tab-delimited
text file. See “Importing Plate Document Setup Table Files” on page B-2 for more information.
Run Setup and Basic Operation 4-3