Qubit
™
4 Fluorometer User Guide
31
Ion Sphere
™
Assay
™
The Ion Sphere
™
assay on the Qubit
™
4 Fluorometer enables quick and easy
assessment of Ion Sphere
™
Particles (ISPs) prior to performing a sequencing run on
the Ion Personal Genome Machine
™
(PGM
™
) Sequencer using the Ion Sphere
™
Quality Control Kit.
All ISPs bind a complimentary, green-fluorescent Alexa Fluor
™
488 dye-labeled
oligonucleotide, whereas only those beads that are template positive bind a
complementary red-fluorescent Alexa Fluor
™
647 dye-labeled oligonucleotide. The
Qubit
™
4 Fluorometer measures the fluorescence emission of each dye conjugate
and a Microsoft Excel
™
file template is used to calculate the percentage of template-
positive ISPs from the ratio of the Alexa Fluor
™
647 dye signal to the
Alexa Fluor
™
488 dye signal. This quality-control step using the Qubit
™
Fluorometer determines the likelihood of a successful sequencing run, ultimately
saving time and expense during subsequent sequencing.
Note: For more information about the Ion Sphere
™
Quality Control Kit
(Cat. No. 4468656), go to thermofisher.com and search for the catalog
number.
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