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Activity Number: 683
Type: Contributed
Date/Time: Thursday, August 8, 2013 : 10:30 AM to 12:20 PM
Sponsor: Biometrics Section
Abstract - #308957
Title: Comparisons of Normalization Methods for Relative Quantification in Real-Time Polymerase Chain Reaction
Author(s): Yi-Ting Hwang*+ and Yu-Hui Su and Harn-Jing Terng and Hsun-Chih Kuo
Companies: National Taipei University and National Taipei University and Advpharma Inc. and Department of Statistics, National Chengchi University
Keywords: oligonucleotide array ; scaling normalization ; invariant set normalization ; quantile normalization ; real-time PCR ; relative quantification
Abstract:

The real-time PCR (real-time polymerase chain reaction) is a common technique for evaluating the gene expression. There are two methods to quantify the real-time PCR gene expression, relative and absolute quantification. Owing to cost and available sources, the relative quantification is the more commonly used method. However, the relative quantification requires a housekeeping gene as an internal control gene to normalize the target gene expression. It has been pointed out the housekeeping gene may be unstable. Hence, it is necessary to discuss how to implement the normalization method for high-density oligonucleotide array to the relative quantification in real-time PCR. Monte Carlo simulations are used to evaluate the performance of three normalizations known in the affymetrix aray to the real-time PCR. Four indices are used to assess the performance. Furthermore, a real data is used to illustrate the feasibility of these normalizations to the real-time PCR. We find that instead of using the housekeeping gene, the scaling normalization is a good choice for relative quantification in real-time PCR.


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