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Proceedings Paper

Multichannel expression analysis of submicrogram total RNA samples without enzymatic amplification using a one-day protocol
Author(s): Robert C. Getts
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Paper Abstract

Typical gene expression array analysis requires relatively large quantities of total or poly A+RNA. Samples prepared by techniques such as laser capture microdissection (LCM) and single cell expression analysis yield relatively little RNA and traditionally require the purification of poly A+ message and subsequent enzymatic amplification before analysis on an array. This type of analysis may be biased to the detection of certain messages, is labor intensive, time consuming and requires considerable expertise for reproducible success. The 3DNATM SubmicroTM expression detection kit has been developed to detect low level expression from a microgram or less of total RNA in one day. The method does not require enzymatic amplification or the direct incorporation of a modified nucleotide during probe synthesis and is simple and easy to use. The 3DNATM detection system is based on patented DNA dendrimers that contain hundreds of fluorescent labels. Signal is generated by the dendrimer after it binds to the cDNA probe (sample) via hybridization of the dendrimer to a capture sequence that is part of the original reverse transcription primer. 50-200 fold improvement of specific signal over noise compared to direct incorporation methods has been demonstrated. The theory and use of the 3DNATM SubmicroTM technology will be discussed for 2,3 and 4 channel analysis.

Paper Details

Date Published: 4 June 2001
PDF: 9 pages
Proc. SPIE 4266, Microarrays: Optical Technologies and Informatics, (4 June 2001); doi: 10.1117/12.427978
Show Author Affiliations
Robert C. Getts, Genisphere Inc. (United States)

Published in SPIE Proceedings Vol. 4266:
Microarrays: Optical Technologies and Informatics
Michael L. Bittner; Yidong Chen; Andreas N. Dorsel; Edward R. Dougherty, Editor(s)

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