JSM 2005 - Toronto

Abstract #303294

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Legend: = Applied Session, = Theme Session, = Presenter
Activity Number: 34
Type: Contributed
Date/Time: Sunday, August 7, 2005 : 2:00 PM to 3:50 PM
Sponsor: Biopharmaceutical Section
Abstract - #303294
Title: Simple Statistical and Graphical Methods in Evaluating Adequacy of Linear Amplification of RNA
Author(s): Yongzheng Ding and Li Xu and Shan Chen and Irene B. Helenowski and Raymond Bergan and Borko Jovanovic*+
Companies: Northwestern University and Northwestern University and Northwestern University and Northwestern University and Northwestern University and Northwestern University
Address: Preventive Medicine, Chicago, IL, 60611, United States
Keywords: RATIO ; COEFFICIENT OF VARIATION ; PROSTATE ; RTPCR ; PC3 ; PC3M
Abstract:

Using LCM to microdissect a population of either pure normal cells or pure cancer cells from the same human prostate gland, we demonstrate the ability to routinely isolate 40 ng of total (high-quality) RNA from a given cell population. We developed a transparent linear amplification protocol that routinely amplifies RNA by more than 1500 fold. We go on to show there is maintenance of gene expression profiles and reproducibility when linear amplified RNA is compared to nonamplified RNA. This was done by first comparing the ratio of gene expression using a 12K gene chip between PC3 and PC3-M prostate cancer cells using 40 mcg of total RNA. Next, the mRNA component (2 ng), from 40 ng of total RNA, were amplified, yielding more than 3.0 mcg of amplified RNA (aRNA). This allowed comparison of PC3/PC3-M gene expression ratios, as measured when nonamplified RNA was used, to PC3/PC3-M gene expression ratios, as measured when aRNA was used.

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Revised March 2005