Qiagen QIAcuity 911000, User Manual

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QIAcuity User Manual 06/2022 8
QIAGEN customers are a major source of information regarding advanced or specialized uses of
our products. This information is helpful to other scientists as well as to the researchers at QIAGEN.
We therefore encourage you to contact us if you have any suggestions about product performance
or new applications and techniques.
For technical assistance and more information, please see our Technical Support Center at
www.qiagen.com/support/technical-support or call one of the QIAGEN Technical Service
Departments or local distributors (see back cover or visit www.qiagen.com ).
Policy statement
It is the policy of QIAGEN to improve products as new techniques and components become
available. QIAGEN reserves the right to change specifications at any time.
To produce useful and appropriate documentation, we appreciate your comments about this user
manual. Please contact QIAGEN Technical Services.
Intended use of the QIAcuity
QIAcuity systems are designed to determine absolute amounts of target DNA in a sample by using
a digital PCR (dPCR) approach.
Digital PCR uses the procedure of end-point PCR, but splits the PCR reaction in many single partitions
in which the template is randomly distributed across all available partitions. After PCR, the target
molecule is detected by measuring the fluorescence – either of sequence specific DNA probes or of
intercalating dyes – in all positive partitions. As the template is distributed randomly, Poisson
statistics can be used to calculate the amount of target DNA per positive partition. The total amount
of target DNA in all partitions of a well is then calculated by multiplying the amount of target DNA
per partition with the number of positive partitions. Calculation of target concentration is determined
by referring back to the volume in all analyzable partitions, i.e., partitions which were filled with
reactions mix. The total number of filled partitions is identified by a fluorescent dye, present in the
reaction mix itself. Absolute quantification by dPCR eliminates the need of standard curves to
determine amounts of target DNA in a given sample.
Aside from absolute quantification, the QIAcuity software provides analysis modules for mutation
detection, genome editing analysis, copy number variation (CNV), and gene expression analysis.