P-MEXP-581 - P-MEXP-581

VIB-MAF Automatic hybridization v1 using Automated Slide Processor (ASP)

A. Reagents:

(1) PolydT35 1 ug/ul (Genset),

(2) Mouse COT-1 DNA (R) 1 mg/ml (Invitrogen, 500 ug, Cat. No. 18440-016),

(3) Human COT-1 DNA (R)1 mg/ml (Invitrogen, 500 ug, Cat. No. 15279-011),

(4) Microarray hybridization buffer (Amersham Pharmacia Biotech, Cat. No. RPK0325),

(5) Formamide (Sigma, Cat. No. F-9037).

Use Mouse COT DNA on Mouse slides, Human COT DNA on Human slides and no COT on Arabidopsis slides.

B. Protocol for automatic hybridization with ASP:

(1) Make 210 ul hybridization mixture per slide with 25% probe, 25% buffer and 50% formamide.

(2) Dissolve the Cy3 and Cy5 sample together in 12 ul of polydT35, 30 ul of Mouse or Human or no (Arabidopsis) COT, water to make a total volume of 52.5 ul, 52.5 ul of Microarray 4x hybridization buffer and 105 ul formamide.

(3) Denature at 96C for 3 minutes.

(4) Keep on ice until hybridization.

(5) Inject samples in ASP.

Download ASP program from
Chamber type, Quantity of label target used, Temperature, Time, Volume
All 12 experiments using protocol P-MEXP-581