A Color-Balanced Spike-In Control for Improved Demultiplexing on XLEAP-SBS Illumina Sequencing Instruments

What is PhiRx?

PhiRx Indexed Control is a dual-indexed control library, made from phiX174 genomic DNA, optimized for Illumina sequencing platforms, particularly two-color systems like XLEAP SBS chemistry on NextSeq 1000/2000 and NovaSeq X.  By improving color balancing for low-plexity libraries PhiRx ensures cleaner, more accurate sequencing results.

Key Features

  • 10 bp i5 and i7 indexes with mixed bases: Provides high base diversity, automatically meeting color balancing requirements on supported platforms. (patent-pending)
  • Color balancing optimization: Specifically designed for the sensitive XLEAP SBS chemistry, enabling as little as 5% spike-in to correct base calling issues in libraries with low-diversity indexes.
  • High diversity for low-plexity samples: Randomized inserts with 45% GC content improve sequencing accuracy for low-diversity libraries.
  • Reliable quality control: Derived from the well-characterized phiX174 genome, providing robust calibration to support sequencing accuracy and performance.

PhiRx Indexed Control reduces the rate of data loss from 53% to < 2%

 

Spike-in Sequencing Control % PhiX Aligned % Undetermined Reads (Excluding PhiX) % One-mismatch
seqWell PhiRx Indexed Control 5.22 1.82 4.15
Illumina PhiX Control V3 6.27 53.25 100.0

 

A 96-plex library prepared using ExpressPlex 2.0 was sequenced on a NextSeq 2000 with 5% spike-in of Illumina PhiX Control V3 or PhiRx from seqWell.

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PhiRx™: A Color-Balanced Spike-In Control for Improving Demultiplexing on XLEAP-SBS Illumina Sequencing Instruments

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