Resolution algorithm
A resolution algorithm, unique when running an HID application, is incorporated
into SeqStudio
™
Data Collection Software v1.2. The purpose of the algorithm is to
align the resolution power traditionally generated with a 3500/3500xL Genetic
Analyzer/POP-4
™
Polymer configuration to the resolution power generated with a
SeqStudio
™
Genetic Analyzer/POP–1
™
Polymer configuration.
About POP
™
polymer matrices
POP
™
polymer matrices separate DNA fragments of a known size range, at a desired
resolution and run time. Polymers are available in different concentrations, which
directly relate to the viscosity and length of the polymer molecule. Different
concentrations are suited to different applications. For example:
• Sequencing analysis applications—Traditionally use POP-6
™
and POP-7
™
Polymers. POP-6
™
and POP-7
™
Polymers are composed of relatively longer
molecules that provide greater DNA-to-polymer interaction and cause DNA
fragments to move through the polymer more slowly. This design results in a
longer run time and the increased resolution that is required for sequencing.
• Fragment analysis applications—Traditionally use POP-4
™
Polymer. Because of
its polymer concentration and molecule size, POP-4
™
Polymer has single basepair
resolving power within the size range for forensic STR amplification kits and
runs faster.
About POP–1
™
Polymer
We are introducing POP–1
™
Polymer for the SeqStudio
™
Genetic Analyzer, which can
be used for all applications on the instrument.
For smaller fragments, POP–1
™
Polymer resolves similarly to POP-7
™
Polymer. For
larger fragments, POP–1
™
Polymer resolves between POP-6
™
and POP-7
™
Polymers.
In addition to providing higher resolution for expected STR fragment peaks, peaks
intrinsic to the amplification process in the n–1/n+1 position from the main allele may
resolve as shoulders with POP–1
™
Polymer. Because of the limited resolving power of
POP-4
™
Polymer, these shoulders were not always visible in our legacy data sets.
The resolution algorithm is applied during data collection. The algorithm maintains
similar resolution for smaller fragments across the 3500/3500xL Genetic Analyzer and
SeqStudio
™
Genetic Analyzer, but allows for the improved resolution for larger
fragments that benefit from the POP–1
™
Polymer.
Appendix D
Create and run an HID plate
Resolution algorithm
D
214
SeqStudio
™
Genetic Analyzer Instrument and Software User Guide