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 experiments using protocol P-MEXP-581: (E-CAGE-198, E-MEXP-186, E-MEXP-2246, E-MEXP-225, E-MEXP-258, E-MEXP-320, E-MEXP-6, E-MEXP-979, E-MEXP-995, E-TABM-67)