Life Technologies
™
| lifetechnologies.com/parkinsons
13
TaqMan
®
SNP Genotyping Assays
A
Figure 4. TaqMan
®
SNP Genotyping Assay. (A) TaqMan
®
SNP Genotyping Assays workflow. Genotyping is performed on the QuantStudio
®
12K Flex Real-Time PCR System using the
TaqMan
®
SNP Genotyping Assay and the 400 base pair amplicon containing the region targeted for TAL editing. Results are analyzed with the TaqMan
®
Genotyper
™
Software. (B) TaqMan
®
SNP Genotyping Assays technology. Discrimination between two SNP alleles (or allelic discrimination) is achieved by selective annealing of two TaqMan
®
probes, where each probe
is specific for one allele and carries a different fluorescent reporter. During PCR amplification, the probe hybridized on the corresponding allele degrades and generates a specific
fluorescent signal. The signal from each reporter is recorded during real-time PCR and plotted on an allelic discrimination plot, where the relative fluorescent signal from allele 1 is plotted
along the x-axis of the allelic discrimination plot and the relative fluorescent signal from allele 2 is plotted along the y-axis. Colonies homozygous for allele 1 will plot along the x-axis.
Colonies homozygous for allele 2 will plot along the y-axis. Heterozygous colonies will plot on a diagonal.
B
Design/order TaqMan
®
Genotyping Assay
Load master mix, assays,
and samples into 96-well plate
Perform real-time PCR
Analysis using TaqMan
®
Genotyper
™
Software
TaqMan
®
SNP Genotyping Assays provide a fast and simple
way to perform single nucleotide polymorphism (SNP)
genotyping analysis using genomic DNA. The workflow is
shown schematically in Figure 4A.
Discrimination between alleles is identified using two different
TaqMan
®
fluorescent reporter probes (Figure 4B).
V
F
F
V
V
F
F
V
Homozygous allele 2 Heterozygous
No-template control Homozygous allele 1
Allelic discrimination plot
Relative fluorescent signal from allele 1
Relative
fluorescent
signal
from
allele
2
Allelic discrimination achieved by selective
annealing of TaqMan
®
probes
Methods
PrOducT BullETIN TaqMan
®
Genotyping Assays
Introduction
Genomic sequencing of a large
number of organisms has produced
a tremendous amount of information
enabling further advances in genomic
analysis, including SNP genotyping.
Applied Biosystems Custom TaqMan
®
SNP Genotyping Assays is a design
service that provides optimized, ready-
to-run assays for any genome, any
organism. These custom assays are
designed, synthesized, formulated,
quality control tested, and delivered in a
convenient, single-tube format. Custom
assays use TaqMan
®
minor groove-
binding (MGB) probe-based assays for
superior allelic discrimination and assay
design flexibility. Simply submit your
target DNA sequence information from
any organism, and we will develop your
ready-to-use SNP genotyping assay.
Custom assays eliminate the time-
consuming manual steps of probe and
primer design, formulation, and testing
to simplify your genomic project plan.
The easy workflow entails adding an
assay plus TaqMan
®
Universal PCR
Master Mix or TaqMan
®
Genotyping
Master Mix with enzyme to your DNA
sample and one-step PCR amplification.
TaqMan
®
Assays enable you to focus
on your strain identification, disease
association, marker selection, quanti-
tative trait loci, linkage mapping, and
other SNP genotyping studies.
Assay design Service
The Custom TaqMan SNP Genotyping
Assays Service provides secure and
confidential access to Applied
Biosystems' assay design expertise.
The service uses the proven Applied
Biosystems Assay Design Pipeline to
create assays designed to your specific-
ations. This versatile resource provides
genotyping assays for any possible SNP,
MNP (up to six base pairs in length),
and InDel (up to six base pairs in length).
Applied Biosystems synthesizes and
tests all assay components in our state-
Custom TaqMan
®
SNP Genotyping Assays
Simplify Your Genomic Projects
•
Any genome, any organism
•
Versatility to detect single nucleotide
polymorphisms (SNPs), insertion/
deletions (InDels), and multi-nucleotide
polymorphisms (MNPs)
•
Proven TaqMan
®
MGB probe-based
assays
•
Made to your specifications
•
One tube, one seamless workflow
