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Quality control of next‐generation sequencing library through an integrative digital microfluidic platform
Author(s) -
Thaitrong Numrin,
Kim Hanyoup,
Renzi Ronald F.,
Bartsch Michael S.,
Meagher Robert J.,
Patel Kamlesh D.
Publication year - 2012
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200441
Subject(s) - microfluidics , capillary action , electrokinetic phenomena , materials science , nanotechnology , analytical chemistry (journal) , computer science , chromatography , chemistry , composite material
We have developed an automated quality control ( QC ) platform for next‐generation sequencing ( NGS ) library characterization by integrating a droplet‐based digital microfluidic ( DMF ) system with a capillary‐based reagent delivery unit and a quantitative CE module. Using an in‐plane capillary– DMF interface, a prepared sample droplet was actuated into position between the ground electrode and the inlet of the separation capillary to complete the circuit for an electrokinetic injection. Using a DNA ladder as an internal standard, the CE module with a compact LIF detector was capable of detecting ds DNA in the range of 5–100 pg/μL, suitable for the amount of DNA required by the I llumina G enome A nalyzer sequencing platform. This DMF ‐ CE platform consumes tenfold less sample volume than the current A gilent B io A nalyzer QC technique, preserving precious sample while providing necessary sensitivity and accuracy for optimal sequencing performance. The ability of this microfluidic system to validate NGS library preparation was demonstrated by examining the effects of limited‐cycle PCR amplification on the size distribution and the yield of I llumina‐compatible libraries, demonstrating that as few as ten cycles of PCR bias the size distribution of the library toward undesirable larger fragments.