Technical Aspects of Microarray Production (updated 12 January, 2000)

Protocols | Array Contents | Troubleshooting | Publications | Technical Experiments

As part of the NSF-funded project: Functional Genomics of Stress Tolerance, we have produced separate microarrays from Arabidopsis, ice plant, rice, maize, and barley. The largest of these arrays (Arabidopsis) contains a total of 18,432 elements from over 9,000 ESTs.

Our current technical focus is on improving reproducibility between hybridization experiments. We are increasing the number of replicate hybridizations and replicate spots for each experiment, and experimenting with automated hybridization stations.

As slide inventories permit, we are willing to provide microarrays to others on a cost-recovery or collaborative basis. See the Galbraith lab homepage for more information on facilities and ongoing microarray projects, and the FAQ for researchers wishing to produce microarrays in collaboration with the Galbraith lab.

Publications on microarray methodology

High density microarrays for gene expression analysis (Deyholos and Galbraith, 2001).

Microarrays for gene discovery and metabolic pathway analysis in plants (Deyholos, Wang and Galbraith, 2001).

Protocols

Large-scale PCR

PCR clean-up using Millipore glass fiber plates

RNA extraction from Arabidopsis or ice plant

RNA extraction from Arabidopsis using TRIZOL

Labeling total RNA

Slide pre-processing

Hybridization and washing

Troubleshooting

Image gallery

Technical experiments

Photobleaching trials

Reproducibility

Database of genes represented on our arrays (links under construction)

Arabidopsis (~9,000 ESTs)

ice plant (~4,000 ESTs)

maize (~4,000 ESTs)

rice (~4,000 ESTs)

barley (~2,000 ESTs)