Applied Biosystems GeneAmp PCR System 9700, User Manual

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B-2 Supplied Methods
About the Methods
Introduction
The instrument supplies you with eight pre-coded methods stored under the user
name <<pe>>.
The eight pre-coded methods are:
♦
AmpliCycle ® Sequencing
♦
AmpliTaq Gold
®
♦
BigDye ™
♦
General PCR
♦
LSM2
♦
Time Release PCR
♦
Touchdown PCR
♦
XL PCR
See the following sections for detailed information about the methods.
AmpliCycle
Sequencing
Cycle sequencing has revolutionized Sanger sequencing of PCR products and other
DNA templates. The AmpliCycle Sequencing protocol is for 3-temperature cycling and
for achieving clean sequence ladders from femtomole amounts of template.
This cycle sequencing process and the benefits of AmpliTaq
®
DNA Polymerase, CS,
are described in the product insert for the AmpliCycle
®
Sequencing Kit
(P/N N808-0175).
AmpliTaq Gold
The AmpliTaq Gold protocol specifies a 5-minute pre-PCR heat step, required for the
activation of AmpliTaq Gold
â
DNA Polymerase. This additional step provides
seamless “hot start” PCR and replaces labor intensive methods such as manual hot
start or wax bead-mediated hot start techniques.
4.0
45:00
72.0
1:00
68.0
0:30
95.0
0:30
95.0
1.00
1 Hld 3 Tmp 25 Cycles 1 Hold
Start Return
Method: AmpliCycle Seq
F1 F2 F3 F4 F5
95.0
5:00
95.0
0:15
55.0
0:15
72.0
0:30
72.0
7:00
2 Holds 3 Tmp 35 Cycles 1 Hld
4.0
F1 F3 F4 F5 F2
Start
Return
Method: AmpliTaq Gold
â