About The 7300/7500/7500 Fast System; About Plus/Minus Assays Using An Ipc; Chapter 1 Introduction - Applied Biosystems 7300 Getting Started Manual

Chapter 1 Introduction

About the 7300/7500/7500 Fast System

About the 7300/7500/7500 Fast System
Description
Plus/Minus Assay

About Plus/Minus Assays Using an IPC

Definition
What Is An IPC?
Notes
2
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Plus/Minus Assay Getting Started Guide
The Applied Biosystems Real Time PCR System (7300/7500/7500 Fast system) uses
fluorescent-based PCR chemistries to provide:
• Quantitative detection of nucleic acid sequence using real-time analysis.
• Qualitative detection of nucleic acid sequence using end-point and dissociation-
curve analysis.
The 7300/7500/7500 Fast system allows you to perform several assay types with plates
in the 96-well format. This guide describes the plus/minus assay, which determines
whether or not a specific target sequence is present in a sample.
For information about the other assay types, refer to the Real-Time PCR Systems
Note:
Chemistry Guide (PN 4378658) and the Online Help for the 7300/7500/7500 Fast
system.
Plus/Minus Assays may be run on a 7500 Fast system using standard reagents;
Note:
Plus/Minus Assays are not supported using Fast reagents and protocols.
A plus/minus assay is an end point assay that determines if a specific target sequence is
present (plus) or not present (minus) in a sample. In an end-point assay, data are
collected at the end of the PCR process.
An IPC is an internal positive control (see TaqMan
Control Reagents kit, PN 4308323) that is used in plus/minus assays to monitor the PCR
process and to ensure that a negative result is not due to failed PCR. The IPC consists of
a template, a primer set, and a dye-labeled (VIC
plate (the IPC is part of the reaction mix, see
page 14).
Plus/Minus assays with an IPC use fluorogenic 5' nuclease chemistry (also known as
"TaqMan probe-based chemistry"). During amplification, the sample target and the IPC
target generate reporter fluorescence signals, so that positive or negative calls may be
made on unknown samples.
®
The SYBR Green I dye chemistry is not supported for plus/minus assays using
Note:
an IPC.
®
Exogenous Internal Positive
®
) probe added to each well of a reaction
"Preparing the PCR Reaction Mix" on