P-MEXP-581 - P-MEXP-581

Type
hybridization
Description
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
Parameters
Chamber type, Quantity of label target used, Temperature, Time, Volume
Links
All 12 experiments using protocol P-MEXP-581