AMX DESIGN XPRESS V 1.5 - PROGRAMMER GUIDE Manuale Utente Scaricare il pdf (Pagina 10)

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December 16, 2004 3:54 pm, GSG_Chapter_4-01.fm

Chapter 1 Before You Begin

Introduction

Primer Express Software 3.0 Getting Started Guide 3

Notes

Primer – A complementary oligonucleotide that initiates amplification of a target region

of DNA. A forward primer anneals to the anti-sense strand. A reverse primer anneals to

the sense strand.

Probe – A short oligonucleotide sequence that anneals specifically to a target sequence

and serves as a fluorescence monitoring system for DNA amplification.

TaqMa n

MGB Probe – An oligonucleotide with a reporter fluorescent dye attached to

the 5′ end and a non-fluorescent quencher attached to the 3′ end. The probe is coupled

with a minor groove binder (MGB), which increases its Tm. When the probe is cleaved

by the DNA polymerase during the PCR reaction, reporter dye fluorescence increases

proportional to the quantity of the target sequence.

TaqMan Probe – An oligonucleotide with a reporter fluorescent dye attached to the 5′

end and a quencher fluorescent dye (usually TAMRA

™

) attached to the 3′ end. When the

probe is cleaved by the DNA polymerase during the PCR reaction, reporter dye

fluorescence increases proportional to the quantity of the target sequence.

Quantification Assay – An assay that determines the relative or absolute quantity of

target sequence within a sample. Relative quantification measures the change in the

expression of the target gene in a test sample, relative to a calibrator sample. Absolute

quantification uses a standard curve to calculate the quantity of an unknown target

sequence.

Sense Strand – In double-stranded DNA, the strand that codes for the RNA that is

translated into proteins. Also referred to as coding, forward, or positive strand. The

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

System

Requirements

The following table lists the hardware and software requirements and recommendations

for installing and using Primer Express version 3.0 software.

Operating Systems Not Supported

• Microsoft

Windows

NT and 2000

• Macintosh

Item Minimum Requirements Recommendations

Computer • Intel

Pentium

III processor

• 540 MHz

•Intel

Pentium IV

processor faster than 2GHz

Monitor • 17-inch monitor

• 800 x 600 pixels resolution

• 19-inch or larger monitor

• 1024 x 768 pixels or higher

pixels resolution

Hard Drives • 256 MB RAM

• 20 MB free hard disk space

• 512 MB RAM

• 10 GB EIDE hard drive

Network Adaptors • 10/100 NIC with RWV (internal)

Printer • Any PC-compatible printer.

Operating System • Windows

XP Professional,

Service Pack 1 or later

•Windows

XP Professional,

Service Pack 1 or later

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