User guide

17
GlobalFiler
™
Express PCR Amplification Kit User Guide
Chapter1 Overview
Instrument and software overview
1
Instrument and software overview
This section provides information about the data collection and analysis software
versions required to run the this kit on specific instruments.
Data collection and
analysis software
The data collection software provides instructions to firmware running on the
instrument and displays instrument status and raw data in real time. As the
instrument measures sample fluorescence with its detection system, the data collection
software collects the data and stores it. The data collection software stores information
about each sample in a sample file (.fsa files for 31xx or 3730 instruments and .hid files
for 3500 instruments), which is then analyzed by the analysis software.
Instrument and
software
compatibility
About
multicomponent
analysis
Applied Biosystems
®
fluorescent multi-color dye technology allows the analysis of
multiple loci, including loci that have alleles with overlapping size ranges. Alleles for
overlapping loci are distinguished by labeling locus-specific primers with different
colored dyes.
Multicomponent analysis is the process that separates the five different fluorescent
dye colors into distinct spectral components. The five dyes used in the GlobalFiler
™
Express Kit to label samples are 6-FAM
™
, VIC
®
, NED
™
, TAZ
™
, and SID
™
dyes. The
sixth dye, LIZ
®
, is used to label the GeneScan
™
600 LIZ
®
Size Standard v2.0.
How
multicomponent
analysis works
Each of these fluorescent dyes emits its maximum fluorescence at a different
wavelength. During data collection on the Applied Biosystems
®
instruments, the
fluorescence signals are separated by a diffraction grating according to their
wavelengths and projected onto a charge-coupled device (CCD) camera in a
predictably spaced pattern. The 6-FAM
™
dye emits at the shortest wavelength and is
displayed as blue, followed by the VIC
®
dye (green), NED
™
dye (yellow), TAZ
™
dye
(red), SID
™
dye (purple), and LIZ
®
dye (orange).
Although each of these dyes emits its maximum fluorescence at a different
wavelength, there is some overlap in the emission spectra between the dyes (Figure 2).
The goal of multicomponent analysis is to correct for spectral overlap.
Instrument
Operating
system
Data collection
software
Additional
software
Analysis
software
3500
†
/
3500xL
†
† We conducted validation studies for the GlobalFiler
™
Express Kit using the 3130xl, 3500, and
3500xL configurations.
Windows
Vista
®
3500 Series Data
Collection Software
v1
HID Updater 3500
DC v2.0
(Part no. 4480670)
GeneMapper
®
ID-X Software
v1.4
•Windows
®
XP
or
•Windows
®
7
Windows
®
7 3500 Series Data
Collection Software
v2
3130/
3130xl
†
Windows
®
7 Data Collection
Software v4
DC v4 6-Dye
Module v1 License
(Contact Life
Technologies)
3730