Benefits of in-situ synthesized microarrays for analysis of gene expression in understudied microorganisms.
Although the genome sequences of many microorganisms are now known, whole-genome DNA microarray platforms consisting of PCR amplicon, or oligonucleotide elements printed onto glass slides have been readily available for only a relatively few, highly studied microorganisms. For those microorganisms more recently cultured or studied by fewer investigators it has been difficult to justify the initial time and expense of developing such array platforms especially if only a limited number of gene expression studies are envisioned. However, in-situ synthesized oligonucleotide (ISO) arrays can be inexpensively fabricated on an 'as needed' basis with a reduced initial investment in time, personnel, resources, and costs. To evaluate the performance of one ISO array platform, gene expression patterns in Geobacter sulfurreducens under nitrogen-fixing conditions were compared with results from quantitative reverse transcriptase PCR (qRT-PCR) and previously published data from a similar experiment using spotted PCR amplicon arrays. There were strong correlations between the results of the ISO arrays and the results from qRT-PCR (r(2)=0.762) and spotted array (r(2)=0.744) analyses. After initial use the ISO arrays could be successfully stripped and reused. The increased flexibility in array design and reusability coupled with a lower initial investment in terms of fabrication time and cost for the ISO arrays suggest that they may be the preferred approach when investigating gene expression in microorganisms, especially when only a few expression studies are required.
Postier
Bradley
B
Department of Microbiology, University of Massachusetts Amherst, MA 01003, USA. bpostier@microbio.umass.edu
Didonato
Raymond
R
Nevin
Kelly P
KP
Liu
Anna
A
Frank
Bryan
B
Lovley
Derek
D
Methe
Barbara A
BA
D001426:Bacterial Proteins / Q000235:genetics; D019295:Computational Biology; D015870:Gene Expression; D045302:Geobacter / Q000235:genetics* / Q000502:physiology; D009586:Nitrogen Fixation; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics / Q000379:methods*; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods
D001426:Bacterial Proteins; D009841:Oligonucleotides