Ratio quantification of gene dosage by Agilent 2100 Bioanalyzer for detection of somatic gene deletions

DOI: 10.4236/abc.2012.21009   PDF   HTML     4,068 Downloads   7,958 Views  

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) < 2%. Then we tested the effect of product concentrations that varied 8-fold, and the ROY was also consistent with CV < 3.5%. Furthermore, we applied this approach to identify six somatic KRAS deletions in non-small cell lung cancer patients, confirming our previous findings. Thus, the Agilent 2100 Bioanalyzer is simple, accurate, quick, and ultimately able to replace conventional gel electrophoresis for the detection of somatic gene deletions.

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Hwu, P. , Liu, Q. , Wang, Y. , Chen, L. , Cheng, Y. , Lin, W. and Yen, Y. (2012) Ratio quantification of gene dosage by Agilent 2100 Bioanalyzer for detection of somatic gene deletions. Advances in Biological Chemistry, 2, 70-75. doi: 10.4236/abc.2012.21009.

Conflicts of Interest

The authors declare no conflicts of interest.

References

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