Thermo Scientific PikoReal, Руководство пользователя

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Introduction to the PikoReal( Real-Time PCR System
Introduction to qPCR
Thermo Fisher Scientific Thermo Scientific PikoReal Real-Time PCR System User Manual 13
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for use with 96-well Piko PCR plates.
The following components form the complete technology platform for
performing real-time PCR based DNA analyses:
●
PikoReal Real-Time PCR System
●
PikoReal Software (see “PikoReal Software” on page 18)
●
UTW® consumables (see “Consumables” on page 18)
●
qPCR reagents (see “Reagents” on page 20).
Quantitative real-time PCR (qPCR) is a method for analyzing DNA by
detecting the amount of PCR product after each PCR cycle by the
incorporation of fluorescent dyes into the reaction. The amplification
and detection of a specific DNA sequence is integrated into a
simultaneous process within a single instrument. The product
accumulation is detected and measured throughout the reaction – in
real-time. The detection is based on monitoring fluorescent molecules
that are incorporated into the amplified products as a dsDNA binding
dye or within labeled target specific primers or probes.
The quantitation is based on correlation between the measured
fluorescence intensity and the amount of DNA formed. Collecting
quantitative data of a known dilution series of the target DNA can be
used to give insight into the PCR reaction kinetics and reaction
efficiency. The gene quantification analysis method can be absolute
(copy number, concentration) or relative (to a known reference gene
and calibrator sample). Examples of qualitative analysis include gene
identification and genotyping, e.g. studying DNA sequence variations,
such as single nucleotide polymorphism.
The basic difference from end-point PCR technique is that in addition
to the qualitative result – detecting the presence of a PCR product, a
quantitative result – the initial amount of the amplified product – can
be determined. qPCR offers many advantages over traditional end-
point PCR methods: high sensitivity over a broad dynamic range,
reproducibility, low contamination risk and the exclusion of post-PCR
manipulations for analysis. It also facilitates high-throughput and
multiplexing applications.
The PikoReal Real-Time PCR System is a thermal cycler with a
fluorescence measuring system available in 24-well and 96-well formats.
It is intended for qPCR technologies and related fluorogenic
chemistries. The PikoReal Real-Time PCR System is intended for
research purposes only.
Use for any other than the above-mentioned purposes is excluded.
Introduction to
qPCR
Intended use