Comparison of commercial probe labeling kits for microarray: towards quality assurance and consistency of reactions.
Microarray technology is readily available to scientists interested in gene expression. Commensurate with this availability is the growing market in accessory products offering convenience but potentially variable performance. Here we evaluate seven commercial kits for probe labeling against a human apoptosis oligonucleotide array. All kits were found to label probes successfully using the manufacturers' instructions. The Stratagene Fairplay Microarray Labeling Kit was the most sensitive, with an overall call rate of 74% and the lowest rate of indeterminant calls for the HEK and HepG2 cell lines. The Invitrogen SuperScript Indirect cDNA Labeling System showed the most reproducible gene expression pattern and the least technical variation, both in terms of signal strength and between replicates on each array. The Promega Pronto! Plus System showed the least dye bias however, a higher level of variation between replicates was observed. Pairwise comparisons revealed that the Promega Pronto! Plus System and Invitrogen SuperScript Indirect cDNA Labeling System had the most similarity in their patterns of gene expression. Results obtained suggest variability in the performance of commercial kits between different manufacturers. This study supports the need to conduct comparative evaluations of commercial microarray probe labeling kits and the need for validation prior to use.
Lynch
Jessica L
JL
Lotterywest State Microarray Facility, School of Medicine and Pharmacology, University of Western Australia, and Centre for Medical Research, Western Australian Institute for Medical Research, Nedlands, WA 6009, Australia.
deSilva
Christopher J S
CJ
Peeva
Violet K
VK
Swanson
Nigel R
NR
D017209:Apoptosis / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D004261:DNA Replication; D018076:DNA, Complementary / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards; D006801:Humans; D046228:Microarray Analysis / Q000379:methods* / Q000592:standards; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods / Q000592:standards; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D018076:DNA, Complementary