Geode H14000 User Manual
4
Introduction to
Crystal Digital PCR™
Purpose of the document
This document provides general information for the use of the Geode for
Crystal Digital PCR™.
The workflow and the different hardware components are described in detail.
Overview of the
Crystal Digital PCR™ Workflow
Crystal Digital PCR
™ is Stilla Technologies’ next-generation technology for the absolute
quantification of nucleic acids.
Using cutting-edge microfluidic innovations, this technology integrates the digital PCR process
in a single consumable (
Fig. 1
). The sample is first flowed through a network of microchannels
and partitioned into a large 2D array of 15,000 to 30,000 individual droplets (Sapphire chips) and
10,000 to 20,000 droplets (Opal chips), also called a droplet crystal. PCR is then performed within
the chips and the crystal is imaged to reveal the droplets that contain amplified targets. The last
step consists of counting the number of these positive droplets to precisely extract the absolute
quantity of nucleic acids.
With
Crystal Digital PCR™, the combination of powerful image analysis and intuitive visual
inspection offers an unmatched level of confidence in the digital PCR measurement, yielding
data you can trust.
Sample loading
Generation of droplet crystal
and PCR
Reading and analysis
Prepare the sample for the
reaction
mix.
Stilla®
recommends the use of the
naica® PCR MIX reagents,
specifically
developed
for
Crystal Digital PCR™. Load
the reaction mix into the wells
of the selected chip, seal with
the provided caps.
Place the prepared chips into the
Geode. Launch the combined
partitioning
and
amplification
program:
droplet
crystals
are
generated from each sample and
PCR amplification is performed
immediately
after
crystal
generation.
After PCR, transfer the chips to
the Prism3 instrument. Set up
the read-out using Crystal
Reader
software
for
data
acquisition of droplet crystals
using up to 3 fluorescent
channels (Blue, Green, and
Red). Image analysis and data
extraction are performed using
the Crystal Miner software.
Figure 1: Overview of
Crystal Digital PCR™ Workflow.
