Purpose
Description
Genotyping experiment
Detects single
nucleotide
polymorphism (SNP)
variants of a
target nucleic acid
sequence.
Genotyping experiments use preformulated TaqMan
™
SNP Genotyping Assays that include
the following components:
• Two sequence-specific primers for amplification of sequences containing the SNP of
interest
• Two allele-specific TaqMan
™
probes for Allele 1 and Allele 2
1.
The software normalizes the fluorescence of the reporter dyes to the fluorescence of
the passive reference dye in each well.
2.
The software plots the normalized reporter dye signal of each sample well on an Allelic
Discrimination Plot, which contrasts the reporter dye intensities of the allele-specific
probes.
3.
The software algorithmically clusters the sample data, and assigns a genotype call to
the samples of each cluster according to its position on the plot.
Presence/absence experiment
Determines the
presence or absence
of a target nucleic
acid sequence in a
sample.
The software calls the target present or absent based on an algorithmically determined call
threshold. (The call threshold is different from the C
t
threshold; the C
t
threshold is not used
to make calls.)
Melt curve experiment
Determines the
melting temperature
(T
m
) of
the amplification
products of a
PCR that used
intercalating dyes.
In the software, melt curve analysis is included in the default run method for any experiment
type that uses intercalating dyes.
1.
The software plots a melt curve based on the fluorescence of the dye with respect to
change in temperature.
2.
Using the melt curve, the software calculates the melting temperature (T
m
).
Chapter 1
Product information
Experiment types
1
20
QuantStudio
™
1 Plus Real-Time PCR System Installation, Use, and Maintenance Guide
