TITLE:
Ratio quantification of gene dosage by Agilent 2100 Bioanalyzer for detection of somatic gene deletions
AUTHORS:
Peter Hwu, Qiang Liu, Yafan Wang, Linling Chen, Yawen Cheng, Wanru Lin, Yun Yen
KEYWORDS:
Electrophoresis; Gene Deletion Detection; Agilent 2100 Bioanalyzer; RD-PCR; NSCLC
JOURNAL NAME:
Advances in Biological Chemistry,
Vol.2 No.1,
February
22,
2012
ABSTRACT: We applied the Agilent 2100 Bioanalyzer, a microfluidics-based electrophoresis instrument, and introduced an accurate and consistent parameter, the relative product yield ratio (ROY), to detect somatic gene deletions in tumor cells. Briefly for such purpose, the Agilent 2100 Bioanalyzer quantified the ROY of a target gene to an endogenous internal control, both of which were initially co-amplified by Robust-Dosage PCR (RD- PCR). Herein, we extensively validated this approach. We first tested the effect of well positions on the Agilent DNA chip. We found that no matter which wells the samples were loaded in, the ROY was consistent with coefficient of variation (CV)