Optimized beadmilling of tissues for high-throughput RNA production and microarray-based analyses.
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.
Hoyt
Peter R
PR
Life Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA. hoytpr@ornl.gov
Doktycz
Mitchel J
MJ
D000818:Animals; D004247:DNA / Q000378:metabolism*; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification
D012313:RNA; D004247:DNA