User guide

12
GlobalFiler
™
Express PCR Amplification Kit User Guide
Chapter1 Overview
Product overview
1
• 3500/3500xL Genetic Analyzer (requires 3500 Data Collection Software v1 or v2
and HID Updater 3500 Data Collection v2)
• 3130/3130xl Genetic Analyzer (requires Data Collection Software v4 and 3130/
3730 Data Collection v4 6-Dye Module v1)
• 3730 Genetic Analyzer (requires Data Collection Software v4 and 3130/3730 Data
Collection v4 6-Dye Module v1)
• GeneMapper
®
ID-X Software v1.4
About the primers
The GlobalFiler
™
Express Kit uses the same primer sequences as the NGM
™
SElect
Express and the Identifiler
®
Direct Kits, which include SNP-specific primers for the
vWA, D16S539, Amel, D2S441, D22S1045, and D8S1179 loci. The GlobalFiler
™
Express
Kit uses the same primer synthesis and purification improvements as the NGM
™
SElect Express and the Identifiler
®
Direct Kits, which enhance the assay signal-to-noise
ratio and simplify the interpretation of results.
The GlobalFiler
™
Express Kit also includes the following primer additions and
modifications:
• Addition of DYS391 and a novel Y indel.
• The TPOX reverse primer has been redesigned to relocate the amplicon into the
higher size range of the multiplex and optimize marker spacing.
• Addition of 8 new SNP-specific primers for the D3S1358, vWA, D18S51, D19S433,
TH01, FGA, D5S818, and SE33 loci. The second degenerate primer was added to
the vWA locus to address two different SNPs found in the primer binding site.
Non-nucleotide linkers are used in primer synthesis for the following loci: D19S433,
vWA, CSF1PO, D2S441, TH01, FGA, and D12S391. For these primers, non-nucleotide
linkers are placed between the primers and the fluorescent dye during oligonucleotide
synthesis (Butler 2005, Grossman et al., 1994). Non-nucleotide linkers enable
reproducible positioning of the alleles to facilitate inter-locus spacing. The
combination of a six-dye fluorescent system and the inclusion of non-nucleotide
linkers allows for simultaneous amplification and efficient separation of all 24 markers
during automated DNA fragment analysis.
Loci amplified by
the kit
Table 1 shows the loci amplified, their chromosomal locations, and the corresponding
fluorescent marker dyes. The GlobalFiler
™
Express Allelic Ladder is used to genotype
the analyzed samples. The alleles contained in the allelic ladder, and the genotype of
the DNA Control 007, are also listed in the table.