Applied Biosystems 7900HT User Manual

DRAFT
September 1, 2004 11:39 am, CH_Basic.fm
Chapter 4 Operating the Instrument
4-10 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
Preparing TaqMan Low Density Arrays for Use
About the Low Density Arrays
The TaqMan Low Density Arrays are research tools for profiling gene expression
using the Comparative C
T
Method of relative quantification. The Low Density Array
evaluates from one to eight cDNA samples or controls generated from human total
RNA in a two-step RT-PCR experiment.
How It Works
The Low Density Array functions as an array of reaction vessels for the
PCR/sequence detection step. Typically, the wells of the Low Density Array contain
Applied Biosystems fluorogenic 5´ nuclease assays (TaqMan
®
reagents; see the note
below) that detect the real-time amplification of user-specified targets. Relative
levels of gene expression are determined from the fluorescence data generated during
PCR using the Applied Biosystems 7900HT Fast Real-Time PCR System Relative
Quantification software.
Note: In this document, the term TaqMan reagents refers to 5´ exonuclease assay
reagents that feature Applied Biosystems TaqMan
®
primers and probes (includes
Applied Biosystems PDARs, TaqMan
®
SNP Genotyping and Gene Expression
Assays, Custom TaqMan
®
SNP Genotyping and Gene Expression Assays, and any
custom TaqMan reagents.)
Low Density
Array
Components
The Low Density Array (Figure 4-5) acts as a vessel for real-time PCR. It consists of
a series of 384 interconnected wells divided into eight sets of assays. Each set
provides a user-specified number of replicates. Each well contains dried Applied
Biosystems TaqMan primers and probes for one mRNA target.
Figure 4-5 The TaqMan Low Density Array
GR2156a
Fill reservoir (1 of 8)
A reservoir for the cDNA sample or
control before it is centrifugally
loaded into the wells
Fill consumable
The segment of the Low Density
Array that contains the eight fill
reservoirs. (After the Low Density
Array is centrifuged and sealed, the
fill consumable is trimmed off.)
Barcode Provides coded access to
Low Density Array configuration
databases

Applied Biosystems 7900HT Specifications

General

Brand	Applied Biosystems
Model	7900HT
Category	Laboratory Equipment
Language	English