Impact of RNA extraction from limited samples on microarray results.
To move microarray technology into the diagnostic realm, the impact of technical parameters, such as sample preparation and RNA extraction, needs to be understood and minimized. We evaluated the impact of two RNA extraction methods, DNase treatment and the amount of hybridized cDNA probe, on the outcome of microarray results. The results for both RNA extraction methods were comparable, although one method resulted in residual DNA that slightly affected the microarray results. As little as one microgram of total RNA could be used to synthesize a cDNA probe and resulted in a gene expression profile that was similar to one produced using 5 micrograms total RNA, even though the overall signal intensity was lower. These experiments illustrate that microarray technology holds great promise for the use of limited clinical samples in the diagnostic setting.
Ojaniemi
Hanna
H
Karolinska Institutet at Huddinge University Hospital, Royal Institute of Technology, Stockholm, Sweden.
Evengard
Birgitta
B
Lee
Daisy R
DR
Unger
Elizabeth R
ER
Vernon
Suzanne D
SD
D016477:Artifacts; D015342:DNA Probes; D020869:Gene Expression Profiling / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000302:isolation & purification*; D015203:Reproducibility of Results; D018401:Sample Size; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*
D015342:DNA Probes; D012313:RNA