AMX DESIGN XPRESS V 1.5 - PROGRAMMER GUIDE Manuel d'utilisateur télécharger pdf (Page 9)

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Chapter 1 Before You Begin

Introduction

2 Primer Express Software 3.0 Getting Started Guide

Notes

Introduction

About Primer

Express

Software

Primer Express software is a primer and probe design tool made specifically for use with

the following instruments:

• Applied Biosystems 7900HT Fast Real-Time PCR System

• Applied Biosystems 7500 Fast Real-Time PCR System

• Applied Biosystems 7500 Real-Time PCR System

• Applied Biosystems 7300 Real-Time PCR System

Primer Express software lets you independently design oligonucleotides (oligos) for

PCR applications using a customized application specific document for each of the

following assay types:

• Absolute/Relative Quantification

• Allelic Discrimination

When using Primer Express software, keep in mind the Applied Biosystems Rapid

Assay Development Guidelines that contain the following important components:

• Design of primers and probes using Primer Express software

• Selection of the appropriate reagent configuration (TaqMan

Universal PCR

Master Mix or SYBR

Green PCR Master Mix)

• Use of universal thermal cycling parameters

• Use of default primer and probe concentrations (or optimizing, if necessary)

IMPORTANT! These components provide a rapid and reliable system for assay design

and optimization only when used in their entirety. Due to the interdependence of many of

the individual components, the system must be adopted as a whole in order to achieve

the highest level of success.

Terms You Need

to Know

Allelic Discrimination Assay – An assay that discriminates between two alleles of

single nucleotide polymorphisms (SNPs). TaqMan

allelic discrimination assays use

two probes specific for the two possible SNP variants.

Anti-Sense Strand – In double-stranded DNA, the strand that does not code for the

RNA, and is not translated into proteins. Also referred to as anti-coding, negative, or

reverse strand. The Primer Express Software designs primers and probes using the sense

strand, not the anti-sense strand.

Document – In the Primer Express Software, a container used to hold sequences,

generate candidate primer and probe designs, and order candidate primer and probe

designs. The four document types available are TaqMan MGB Quantification, TaqMan

Quantification, TaqMan MGB Allelic Discrimination, and TaqMan Allelic

Discrimination.

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