Applied Biosystems GeneAmp PCR System 9700, User Manual

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B-4 Supplied Methods
LMS2
The GeneAmp PCR System 9700 software includes a pre-coded LMS2 method for
ABI P
RISM®
Linkage Mapping Set Version 2 (LMS2) thermal cycling. Linkage Mapping
Set 2 employs over 400 fluorescent-labeled PCR primer pairs for analysis of select
microsatalite loci from the Généthon human linkage map.
1,2,3 The following figures
illustrate the thermal cycling profile for the LMS2 method.
This supplied method consists of an initial hold at 95.0 °C, two sets of three
temperature cycles, followed by two additional holds at 72.0 °C and 4.0 °C.
See the ABI P
RISM
Linkage Mapping Set Version 2 User’s Manual (P/N 904999) for
comprehensive information on PCR amplification conditions, electrophoresis
conditions, detection, and data analysis.
Time Release PCR
The Time Release PCR method is designed for use with AmpliTaq Gold DNA
Polymerase. The enzyme is activated more slowly than with the AmpliTaq Gold
method. Here the pre-PCR hold is only 1 minute, and the number of cycles is
increased to 40.
1. Weissenbach, J. et al. 1992. A second-generation linkage map of the human genome. Nature
359:794–801.
2. Gyapay, G., et al. 1994. Généthon Human Genetic Linkage Map. Nature Genet. 7:246–339.
3. Dib, C., et al. 1996. Nature 380: 152–154.
3 Tmp 20 Cycles 3 Tmp 10 Cycles
89.0
0:15
Start Return
Method: LMS2
F1 F2 F3 F4 F5
95.0
12:00
94.0
0:15
55.0
0:15
72.0
0:30
55.0
0:15
72.0
0:30
1 Hld
3 Tmp 20 Cycles 3 Tmp x 10
89.0
0:15
Start Return
Method: LMS2
F1 F2 F3 F4 F5
55.0
0:15
72.0
0:30
55.0
0:15
72.0
0:30
72.0
10:00
4.0
×
2 Holds
4.0
72.0
0:30
55.0
0:15
95.0
0:15
95.0
1.00
1 Hld 3 Tmp 40 Cycles 2 Holds
Start Return
Method: Time Release PCR
F1 F2 F3 F4 F5
72.0
7:00