Geode H14000 User Manual
12
provides a certification documenting and guaranteeing adequate naica® system installation and
validation. The certification is required to release the naica® system for customer operation.
g) Materials required but NOT supplied
i.
Crystal Digital PCR™ recommended PCR mastermixes
Depending on the application, Stilla Technologies recommends the use of naica® multiplex PCR
MIX for fluorescently labeled probe-based digital PCR assays (TaqMan®) or naica® PCR MIX
for EvaGreen® dye-based digital PCR assays for
optimal results with Crystal Digital PCR™ for
DNA amplification. These reagents are available at 5X and 10X concentrations and Stilla
Technologies provides fast cycle or conventional PCR cycling protocols for initial protocol
development.
More information is available at:
https://www.stillatechnologies.com/naica-system-dpcr-mixes/
For RT-PCR applications, Stilla Technologies recommends the use of qScript
TM
XLT One-Step
RT-qPCR ToughMix® (Quantabio).
As an alternative, the following PCR mastermixes are validated for the naica® system:
PerfeCTa® MultiPlex qPCR ToughMix®, the PerfeCTa® qPCR ToughMix® UNG, and
PerfeCTa® qPCR ToughMix® (Quantabio). When using cDNA, we recommend using the
qScript® Flex cDNA Synthesis kit (Quantabio). For further information and instructions, please
refer
to
the
Technical
Resources
webpage
available
at
https://www.stillatechnologies.com/technical-resources/naica-system-prism3/
.
Any other enzymatic mix needs to be evaluated by the user before performing experiments, as
Stilla Technologies cannot guarantee the compatibility of any other enzymatic mix with the
naica® system.
h)
Crystal Digital PCR™ recommended detection system
For the detection and quantification of the amount of target in the sample, a fluorescent signal is
measured. The Crystal Digital PCR
TM
technology allows the use of fluorescently labeled
hydrolysis probes (TaqMan®) as well as the non-specific fluorescent intercalating dye
EvaGreen®.
Note: SYBR® Green is not compatible with the naica
®
system.
i.
Fluorescently labeled probes
The Crystal Digital PCR
TM
technology allows the detection of up to 3 different targets in the same
sample using 3 different fluorophores, depending on the configuration.
A fluorophore is a fluorescent chemical compound that absorbs light energy of a specific
wavelength and emits light at a longer wavelength. They are used in many applications and are
characterized notably by their excitation/emission spectra. In order to be detected by the Prism3
instrument, a fluorophore excitation spectrum must overlap with that of the LED of a given
detection channel, and its emission spectrum must fall within the bandpass of the tri-band filter
used for acquisition (
Table 1
).
Table 1: The Prism3 optical specifications and examples of compatible fluorophores.
Blue Channel
Green Channel
Red Channel
Excitation range
415-480 nm
530-550 nm
630-640 nm
Emission range
495-515 nm
560-610 nm
655-720 nm
Compatible fluorophores
(reporter)
FAM, Fluorescein
Cy®3, HEX
Alexa Fluor® 647,
Cy®5
