Production of bulk amounts of universal RNA for DNA microarrays.
In DNA microarray technology, repeatability and reliability are very important to compare multiple RNA samplesfrom different experiments. The application of common or universal RNA as a standard control equalizes the differences in hybridization parameters and array variations. For this purpose, high-quality reference RNA is necessary in bulk amounts. A novel approach was developed to get milligrams of sense or antisense RNA, starting from micrograms of pooled total RNA from different cell lines, tissues, or organisms. This method is inexpensive and allows further labeling procedures using poly(dT) or random oligomers as primers. In addition, amplified, sense reference RNA is suitable for standard labeling protocols, while the antisense reference RNA can be used with antisense RNA from the linear sample amplification method. Here we produced universal RNA for human, rat, and alfalfa and demonstrated the quality using specific cDNA microarrays.
Puskás
L G
LG
DNA-chip Laboratory, Biological Research Center, Hungarian Academy of Sciences, Szeged. pusi@nucleus.szbk.u-szeged.hu
Zvara
A
A
Hackler
L
L
Micsik
T
T
van Hummelen
P
P
D000818:Animals; D001483:Base Sequence; D006801:Humans; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification*; D051381:Rats
D012313:RNA