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Library preparation with QIAseq Human Comprehensive Cancer panel

SL So Ngo Lam
YZ Ying Chun Zhou
YC Yee Man Chan
CF Ching Man Foo
PL Po Yi Lee
WM Wing Yeung Mok
WW Wing Sum Wong
YF Yan Yee Fung
KW Kit Yee Wong
JH Jun Yuan Huang
CC Chun Kin Chow
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The kit was purchased from QIAgen (Hilden, Germany). Sequencing libraries were prepared according to the manufacturer’s instructions. Briefly, 15 ng of plasma cfDNA fragments were end-repaired, A-tailed, ligated with UMI-barcoded adaptors. The adaptor-ligated libraries were target enriched with PCR using a panel of loci specific primers (6 cycles). The target enriched libraries were further amplified for 21 cycles with PCR and were size selected for an average fragment size of 350 bp. The library profile was analyzed with the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) and quantified using QuBit dsDNA HS Assay kit with QuBit 3.0 fluorimeter (Thermo Fisher Scientific, San Jose, CA) and qPCR with QIAseq Library Quant Assay Kit (QIAgen, Hilden, Germany).

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