Applied Biosystems 7900HT User Manual

DRAFT
September 1, 2004 11:39 am, CH_Real-Time.fm
Chapter 6 Analyzing Real-Time Data
6-12 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
c. Do one of the following. Select the:
Automatic Baseline option button to have the software calculate a baseline
for the selected detector(s).
Manual Baseline option button, and enter Start and End cycle values to
apply to the selected detector(s).
Manual Baseline option button, and leave the Start and End cycles fields
empty. (You will need to set the values manually as explained on page 6-46.)
4. If necessary, repeat steps 2 through 3 for any additional detectors.
IMPORTANT! The threshold and baseline values set in step 3 on page 6-11 are
detector specific, and must be set for each individual detector used on in the
plate document.
5. Click to accept the analysis settings and exit.
6. Analyze the data as explained below.
Analyzing the
Run
After you have configured the analysis settings, you can analyze the run data. During
the analysis, the software mathematically transforms the raw data to establish a
comparative relationship between the spectral changes in the passive reference dye
and those of the reporter dye. Based on that comparison, the software calculates a
cycle threshold (C
T
) for each reaction (standard and unknown). The software then
generates a standard curve for the run by plotting the standard samples on a graph of
C
T
versus initial copy number.
Note: See Appendix D, “Theory of Operation,” for a detailed description of the SDS
software mathematical transformation of real-time run data.
1. Click (or select Analysis > Analyze).
The SDS software analyzes the run data and displays the results in the Results
tab.
2. Choose from the following:
• If using the AutoC
T
algorithm to set the baseline and threshold values for
the detectors on the plate, verify that the software has called the settings
correctly for each detector as described on page 6-46,
• If you choose to set the baseline and threshold values manually, set them
for each detector on the plate as explained on page 6-46.
3. If you choose to remove outliers manually, visualize and eliminate outliers from
the analyzed run data as explained on page 6-46.

Applied Biosystems 7900HT Specifications

General

Brand	Applied Biosystems
Model	7900HT
Category	Laboratory Equipment
Language	English