Optimized beadmilling of tissues for high-throughput RNA production and microarray-based analyses

Peter R. Hoyt; Mitchel J. Doktycz

doi:10.1016/j.ab.2004.04.025

Optimized beadmilling of tissues for high-throughput RNA production and microarray-based analyses

Peter R. Hoyt, Mitchel J. Doktycz

Biosciences Division

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

The preparation of RNA samples has become the rate-limiting step when performing genome-scale analyses by DNA microarrays. Methods to improve throughput of RNA isolation from tissues are needed. The effects of bead size and composition for disrupting mouse tissues have been evaluated in small centrifuge tubes and optimized for RNA production. The resulting process is inexpensive, resistant to cross-contamination, and amenable to robotic processing. After optimization, very-high-quality RNA can be produced. Comparisons between RNAs isolated by beadmilling (followed by solid-phase purification) and those by conventional isolation processes show that RNA produced by beadmilling is suitable for microarray analyses. Parallel implementation of beadmilling will enable a high-throughput tissue-to-RNA processing system for large-scale microarray analyses.

Original language	English
Pages (from-to)	100-108
Number of pages	9
Journal	Analytical Biochemistry
Volume	332
Issue number	1
DOIs	https://doi.org/10.1016/j.ab.2004.04.025
State	Published - Sep 1 2004

Funding

This research was sponsored by the Office of Biological and Environmental Research, U.S. Department of Energy through Oak Ridge National Laboratory (ORNL) Directors Research and Development Funds and ORNL Seed Money Program Funds. ORNL is managed by UT-Battelle, LLC, for the U.S. Department of Energy under Contract No. DE-AC05-00OR22725.

Keywords

Beadmilling
DNA array
High throughput
Microarray
RNA
Sample preparation

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10.1016/j.ab.2004.04.025

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abstract = "The preparation of RNA samples has become the rate-limiting step when performing genome-scale analyses by DNA microarrays. Methods to improve throughput of RNA isolation from tissues are needed. The effects of bead size and composition for disrupting mouse tissues have been evaluated in small centrifuge tubes and optimized for RNA production. The resulting process is inexpensive, resistant to cross-contamination, and amenable to robotic processing. After optimization, very-high-quality RNA can be produced. Comparisons between RNAs isolated by beadmilling (followed by solid-phase purification) and those by conventional isolation processes show that RNA produced by beadmilling is suitable for microarray analyses. Parallel implementation of beadmilling will enable a high-throughput tissue-to-RNA processing system for large-scale microarray analyses.",

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AB - The preparation of RNA samples has become the rate-limiting step when performing genome-scale analyses by DNA microarrays. Methods to improve throughput of RNA isolation from tissues are needed. The effects of bead size and composition for disrupting mouse tissues have been evaluated in small centrifuge tubes and optimized for RNA production. The resulting process is inexpensive, resistant to cross-contamination, and amenable to robotic processing. After optimization, very-high-quality RNA can be produced. Comparisons between RNAs isolated by beadmilling (followed by solid-phase purification) and those by conventional isolation processes show that RNA produced by beadmilling is suitable for microarray analyses. Parallel implementation of beadmilling will enable a high-throughput tissue-to-RNA processing system for large-scale microarray analyses.

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KW - High throughput

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Optimized beadmilling of tissues for high-throughput RNA production and microarray-based analyses

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