Protocols - Microarray Hybridization
Array hybridization is performed on the Genomic Solutions Automated Hybridization Station. This system allows us to vary temperature, hybridization time, and wash conditions of the array. Up to twelve arrays can be completed at one time with six different hybridization conditions. The facility will optimize conditions for your array to produce highest stringency hybridization with the most favorable signal to noise ratio. Currently, our full length cDNA arrays are run under the following conditions; however, we have modified this protocol for some arrays taking into account average probe length, GC content, and amount of labeled material.
1. Inject probe at 68 °C
2. Hybridize at 47 °C for 14-16hrs
3. Drain hybridization solution
4. Low stringency wash at 50°C for 20sec x 2
5. Drain
6. Medium stringency wash at 42°C for 20sec x 2
7. Drain
8. High stringency wash at 42°C for 20sec x 2
9. Drain
10. Remove slides from cassette, dunk in fresh milli-Q water 5 times, and spin dry in centrifuge
