Multichannel expression analysis of submicrogram total RNA samples without enzymatic amplification using a one-day protocol

Robert C. Getts

doi:10.1117/12.427978

4 June 2001 Multichannel expression analysis of submicrogram total RNA samples without enzymatic amplification using a one-day protocol

Robert C. Getts

Author Affiliations +

Proceedings Volume 4266, Microarrays: Optical Technologies and Informatics; (2001) https://doi.org/10.1117/12.427978
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

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 Submicro^TM 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 3DNA^TM 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 Submicro^TM technology will be discussed for 2,3 and 4 channel analysis.

Citation Download Citation

Robert C. Getts "Multichannel expression analysis of submicrogram total RNA samples without enzymatic amplification using a one-day protocol", Proc. SPIE 4266, Microarrays: Optical Technologies and Informatics, (4 June 2001); https://doi.org/10.1117/12.427978

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
9 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Dendrimers

Statistical analysis

Lung

Visualization

Fluorescent markers

Target detection

Detector development

Show All Keywords

Keywords/Phrases

Search In:

Publication Years